J Mol Cell Cardiol 33, 835–849 (2001)

doi:10.1006/jmcc.2000.1317, available online at http://www.idealibrary.com on

Basic Cardiac Electrophysiology

IKr: The hERG Channel

Gea-Ny Tseng
Department of Physiology, Virginia Commonwealth University, Richmond, VA 23298, USA
(Received 29 November 2000, accepted 6 December 2000, published electronically 24 January 2001)

G.-N. T. IKr: The hERG Channel. Journal of Molecular and Cellular Cardiology (2001) 33, 835–849. The rapid
delayed rectifier (IKr) channel is important for cardiac action potential repolarization. Suppressing IKr function,
due to either genetic defects in its pore-forming subunit (hERG) or adverse drug effects, can lead to long-QT
(LQT) syndrome that carries increased risk of life-threatening arrhythmias. The implication of IKr in cardiac
arrhythmias and in anti-arrhythmic/pro-arrhythmic actions of drugs has driven intensive research interests in
its structure–function relationship, the linkage between LQT-associated mutations and changes in channel
function, and the mechanism of drug actions. This review will cover the following topics: (1) heterogeneous
contribution of IKr to action potential repolarization in the heart, (2) structure–function relationship of IKr/hERG
channels, (3) role of regulatory
subunits in IKr/hERG channel function, (4) structural basis for the unique
pharmacological properties of IKr/hERG channels, and (5) IKr/hERG channel modulation by changes in cellular
milieu under physiological and pathological conditions of the heart. It is anticipated that further advances in our
understanding of IKr/hERG, particularly in the areas of roles of different ( and
) subunits in native IKr function,
alterations in IKr function in diseased hearts, and the 3-dimensional structure of the IKr/hERG pore based on
homology modeling using the KcsA model, will help us better define the role of IKr in arrhythmias and design
therapeutic agents that can increase IKr and are useful for LQT syndrome.  2001 Academic Press

K W: IKr, Human ether-a-go-go related gene, Long-QT syndrome, Class III anti-arrhythmic drugs.

Introduction the molecular basis of IKr channel function in the

The rapid component of delayed rectifier (IKr) con-
tributes to phase 3 repolarization in cardiac action
potentials.1,2 Genetic defects in the major pore-form-
ing subunit (hERG)3 or in the putative regulatory (
)
subunit (MiRP1)4 of IKr channels are associated with
congenital (LQT2 and LQT6) and acquired long-QT
syndrome,5 that carries increased risk of life-threat-
ening cardiac arrhythmias. IKr is the target of a group
of potent and specific class III anti-arrhythmic drugs,
methanesulfonanilides.6,7 It can also be blocked by a
wide range of pharmacological agents with different
chemical structures. The implication of IKr in cardiac
arrhythmias and in anti-arrhythmic or pro-arrhy-
thmic actions of drugs highlights the importance of
understanding its role in action potential re-
polarization, its structure–function relationship, and
the mechanism of drug actions on IKr. Tremendous
insights into these issues have been generated by the
intensive research efforts during the past few years.
These have led to excellent recent reviews discussing
Please address all correspondence to: Gea-Ny Tseng, PhD, Department of Pathology, Virginia Commonwealth University, 1101 E.
Marshall Street, Richmond, VA 23298, USA; Phone: 804-827-0811; Fax: 804-828-7382; E-mail: gtseng@hsc.vcu.edu
heart,8 the biophysical properties of IKr/hERG gating,
9
LQT-associated mutations in hERG,10,11 and the
mechanisms for congenital and acquired LQT syn-
drome.12,13
This minireview will begin with a discussion
of the heterogeneous contribution of IKr to action
potential repolarization in the heart. This topic
bears fundamental importance in the attempt to
understand the linkage between defects in IKr chan-
nel function (either genetic or acquired) and
arrhythmogenesis. This is followed by a summary
of current knowledge about the structure-function
relationship of IKr/hERG channels. The emphasis of
this discussion is on aspects that are relevant for
understanding structural determinants of IKr con-
tribution to action potential repolarization, and of
IKr blockade by drugs. The two following sections
are about new concepts that have emerged since
previous reviews (‘‘Role of regulatory
subunits in
IKr/hERG channel function’’, ‘‘structural basis for
the unique pharmacological properties of IKr/hERG

0022–2828/01/050835+15 $35.00/0  2001 Academic Press

836
channels’’). The last section is a brief discussion of
IKr/hERG channel modulation by changes in cellular
milieu under physiological and pathological con-
ditions of the heart. This information is important
for the understanding of arrhythmogenesis in dis-
eased hearts, and the design of therapeutic in-
terventions useful for these conditions.
Heterogeneous Contribution of IKr to
Action Potential Repolarization in the
Heart
IKr was first identified in guinea pig ventricular
myocytes.6 Subsequently, currents with similar gat-
ing and pharmacological properties have been de-
scribed for various cardiac cell types (atria, nodal,
Purkinje and ventricular cells) from a wide variety
of animal species: guinea pig,14 ferret,15 mouse,16–18
rat,19,20 cat,21,22 rabbit,23–25 and dog.26–29 IKr has also
been described for human atrial30 and ventricular31
myocytes. IKr channels contribute outward currents
to action potential repolarization at voltages neg-
ative to 0 mV (phase 3 repolarization). However,
the role of IKr in action potential repolarization is
subject to modulation by ‘‘intrinsic’’ and ‘‘extrinsic’’
factors.
Intrinsic factors
These include two factors. The first is the expression
level of channel subunits. A detailed study of
regional localization of ERG protein and mRNA in
ferret heart32 revealed that ERG expression is most
abundant in the epicardial cell layers throughout
most of the ventricles, except at the base. In the
atria, the ERG mRNA and protein are most abund-
ant in the medial right atrium, especially in the
trabeculae and the crista terminalis of the right
atrial appendage. The distribution pattern of ERG
in ferret ventricles is echoed in functional studies
of IKr current density in other species. For example,
in rabbit left ventricle, IKr current density is larger
in myocytes from the apex than those from the
base.33 In the left ventricles of guinea pig and
cat, IKr current density is significantly higher in
myocytes from the epicardial surface than those
from the endocardial surface.34,35 Pond et al. used
the immunoblot and voltage clamp techniques to
show that both ERG protein level and IKr current
density are higher in rat atria than in rat vent-
ricles.20 However, in human heart hERG expression
is higher in ventricles than in atria.20 Therefore, IKr
G.-N. Tseng
expression is heterogeneous in the heart, and the
expression pattern shows some species variations.
The second intrinsic factor that can influence IKr
contribution to action potential repolarization is its
gating kinetics. The IKr channels are activated dur-
ing the action potential’s upstroke and plateau
phase (phase 2). At voltages positive to 0 mV, a fast
inactivation process of IKr channels greatly limits the
amount of outward current through the channels
(inward rectification). As the membrane voltage
repolarizes to below 0 mV (phase 3), IKr channels
recover from inactivation and reenter the open
state, leading to an outward ‘‘resurgent’’ current
that facilitates further repolarization.1,2 The out-
ward IKr current subsides toward the end of action
potential due to channel deactivation and a decrease
in driving force for outward currents. The voltage-
and time-dependence of IKr activation will influence
the degree of channel activation during phase 2,
while the processes of recovery from inactivation
and subsequent deactivation determine the amount
of outward IKr current during phase 3.
The voltage-dependence of IKr activation, as well
as the processes of inactivation and recovery from
inactivation, is similar in myocytes from different
regions of the heart and from different species.6,14,22,
27,28,30,31,36
Therefore, channel activation has a
threshold of −40 to −30 mV and reaches a plateau
at +20 to +30 mV. Outward IKr current during de-
polarization test pulses peaks at 0 to +10 mV and
inwardly rectifies at more positive voltages. On the
other hand, there are marked differences in the kin-
etics of IKr activation and deactivation in myocytes
from different species. Table 1 lists some of the  val-
ues of IKr activation and deactivation available in
the literature. To minimize differences arising from
different recording conditions, all the studies in-
cluded in Table 1 had comparable recording con-
ditions [35–37°C, IKr measured as drug (E-4031 or
dofetilide)-sensitive currents, [K+]o=4–6 m, ex-
ternal divalent cation concentration=2–3 m]. For
both activation and deactivation rates, guinea pig IKr
channels are relatively fast, rabbit, canine and cat IKr
channels are slow, and human IKr channels are in
between. Gintant29 used voltage clamp waveforms
simulating action potentials to characterize the out-
ward IKr transients during repolarization in canine
ventricular, atrial and Purkinje myocytes. In all three
cell types, there was a dispersion of voltages at which
the peak IKr transients occurred. Since the timing of
outward IKr transients during repolarization depends
on the rate of recovery from inactivation (fast and
invariant among cells) and the rate of deactivation,
these data suggest that there are regional variations
in the kinetics of IKr deactivation in canine heart.
Table 1 Kinetics of activation and deactivation of native IKr in adult cardiac myocytes and hERG channels in heterologous systems

Activation  (ms) Deactivation  (ms)

0 mV +10 mV +20 mV +30 mV +40 mV +50 mV +60 mV −40 mV −50 mV −60 mV Reference

Native Guinea pig ventricle 15 140 39 6

IKr
Guinea pig atrium 40 15 119 30 14
Rabbit ventricle 400 200 600 250 36
Canine ventricle 600 50 500 200 27
Cat ventricle 1511±382 201.2±40.1 80.2±19.2 250 125 22
Human ventricle 192±53 31
Human atrium 200 100 234±24 30
hERG in HEK 293 cells 105±15 200 149±27 100 2
IKr: The hERG Channel

channel
in CHO cells 58.6±1.8 29.2±5.4 1

All experiments were done under comparable conditions: 35–37°C, [K+]o=4–6 m, extracellular divalent cation concentration 2–3 m. IKr was measured as drug (E-4031 or dofetilide)-
sensitive currents. The activation  was determined by fitting single exponential function to the test pulse currents during depolarization or to the envelope of tail currents after depolarization
pulses of different durations. The deactivation  was determined by fitting single or double exponential function to tail currents at specified voltages. In the latter case, the  value of the fast
component is listed here.
837
838 G.-N. Tseng
These differences in IKr kinetics may be due to varying
gating properties of ERG isoforms, or due to vari-
ations in IKr channel subunit composition.4,37,38
An E-4031 (5 )-sensitive current with novel
gating properties has been described for canine
subendocardial Purkinje myocytes at 35–36°C in
normal Tyrode’s solution.39 This E-4031-sensitive
current does not inwardly rectify even at voltages
up to +60 mV, and has extremely fast deactivation
kinetics (at −40 mV). Furthermore, its time course
of activation depends on the prepulse potential
(Vc): at Vc −40 mV the activation appears to be
instantaneous, while at Vc −90 mV there is a
distinct phase of time-dependent activation with
>40–50 ms (at +20 mV). These gating char-
acteristics are similar to those of ether-a-go-go
(EAG) channels.40 These data suggest that canine
subendocardial Purkinje myocytes may express an
ERG isoform with gating kinetics similar to those
of a related gene product (EAG). The unique gating
kinetics suggests that this current may play an
important role in determining the action potential
configuration and duration in these Purkinje
myocytes.39
Extrinisic factors
These include two factors. The first is the function
and expression of other currents and thus the
configuration and rate of action potentials. At
35–37°C and +20 mV, IKr activation has a  value
ranging from 15 to 200 ms (Table 1). Therefore,
action potentials with a positive and long-lasting
plateau phase (e.g. ventricular action potentials of
guinea pig) will allow IKr to activate to a higher
level than action potentials with a negative or
short plateau phase (atrial action potentials and
ventricular action potentials of adult rat and
mouse). For guinea pig IKr that activates and de-
activates rapidly (Table 1), its role in action potential
repolarization will not be rate- (or use-) dependent.41
On the other hand, for IKr channels that activate
and deactivate more slowly (e.g. cat IKr, Table 1),
its contribution to action potential repolarization is
likely to be use-dependent. This will be especially
the case under abnormal conditions, when the
action potential duration is shortened (reducing
the degree of IKr activation during a single action
potential), and the diastolic potential is partially
depolarized (slowing IKr deactivation and facilitating
accumulation of IKr activation during successive
action potentials). Indeed, in adult cat ventricular
myocytes, the IKr tail current amplitude shows a
use-dependence at a holding voltage of −40 mV,
but not at −80 mV.22
The second extrinsic factor that can affect IKr
contribution to action potential repolarization is
modulation by cellular milieu under physiological
or pathological conditions. Local [K]o is an im-
portant determinant of IKr current amplitude and
deactivation kinetics.42–44 Local changes in pHo45 or

-adrenergic stimulation46,47 may have important
effects on IKr deactivation kinetics. Pathological con-
ditions of the heart can induce long-term changes
in IKr subunit expression and IKr channel function,
that may have differential effects on IKr contribution
to action potential repolarization in different regions
of the heart.39,48,49
Therefore, the expression level and gating kinetics
of IKr channels (intrinsic factors) are heterogeneous
in the heart. IKr contribution to action potential
repolarization is further subject to modulation by
regional variations in action potential configuration
and changes in cellular milieu (extrinsic factors).
The heterogeneous contribution of IKr to action
potential repolarization is important for normal
heart function. However, heterogeneous changes in
IKr amplitude and/or gating kinetics under abnormal
conditions48,49 may enhance the dispersion of re-
polarization and refractoriness in the heart, setting
the stage for reentrant arrhythmias.
Structure–function relationship of the hERG channel
The hERG channel has the same body plan as that
of other voltage-gated ion channels50–52: each hERG
subunit has six transmembrane -helices, with the
fourth one (S4) carrying seven positive charges dis-
tributed at every third or fourth position, and a re-
entrant ‘‘pore-loop’’ between the fifth and sixth
transmembrane helices that contains a slightly
modified version of the ‘‘signature sequence’’ of K-
selective pores (Fig. 1).53,54 Based on structure–
function analysis of other voltage-gated K (Kv) chan-
nels, it is inferred that each functional hERG channel
is composed of four subunits surrounding a central
aqueous pore, with the outer one-third lined by the
pore-loops from the four subunits forming a narrow
‘‘selectivity filter’’, and the inner two-thirds lined by
the carboxyl-halves of the S6 domains that form the
inner vestibule of the pore.54,55
The following discussion of the structure–
function relationship of hERG channel emphasizes
aspects that are relevant for understanding the
structural determinants of IKr contribution to action
potential repolarization, the molecular basis for
 IKr: The hERG Channel 839

652 656
↓ ↓
hERG ALYFTFSSLTSVGFGNVSPNTNSEKIFSICVMLIGSLMYASIFGNVSAIIQRLY
KcsA ALWWSVETATTVGYGDLYPVTLWGRLVAVVVMVAGITSFGLVTAALATWFVGRE
Shaker AFWWAVVTMTTVGYGDMTPVGFWGKIVGSLCVIAGVLTIALPVPVIVSNFNYFY
Kv1 AFWWAVVSMTTVGYGDMYPVTIGGKIVGSLCAIAGVLTIALPVPVIVSNFNYFY
Kv2 SFWWATITMTTVGYGDIYPKTLLGKIVGGLCCIAGVLVIALPIPIIVNNFSEFY
Kv3 GFWWAVVTMTTLGYGDMYPQTWSGMLVGALCALAGVLTIAMPVPVIVNNFGMYY
Kv4 SFWYTIVTMTTLGYGDMVPKTIAGKIFGSICSLSGVLVIALPVPVIVTNFSRIY
KvLQT1 ALWWGVVTVTTIGYGDKVPQTWVGKTIASCFSVFAISFFALPAGILGSGFALKV
P-loop S6 helix
Figure 1 Alignment of amino acid sequences of the pore (P)-loop and S6 helix regions. The residues discussed in the
text are highlighted in bold italics. Numbers on top denote the two unique aromatic amino acids in hERG’s S6 that
are important for drug binding.

defects in LQT-associated mutations, and the mech-
anism of IKr blockade by drugs.
Activation and deactivation gating
Studies on other Kv channels have suggested that
the activation ‘‘gate’’ is formed by the S4–S5 linkers
(cytoplasmic loops connecting the S4 and S5 trans-
membrane -helices) and the carboxyl half of the
S6 domains (C-half of S6).56–58 This is probably also
the case for the hERG channel. Therefore, point
mutations in the S4-S5 linker and in the C-half of
S6 have profound effects on the voltage-dependence
and kinetics of channel activation and de-
activation.55,59,60 Such a structural arrangement
places the activation gate close to the cytoplasmic
entrance of the pore. This can explain why hERG
pore blockers that work from the cytoplasmic side
of the membrane (e.g. methanesulfonanilides) gain
access to the binding site within the pore only when
the activation gate is in the open position (open
state blockade).7,61
One important determinant of IKr ’s contribution
to action potential repolarization is its uniquely
slow deactivation process. Therefore, some LQT2-
associated mutations reduce outward currents
through the IKr channels by accelerating channel
deactivation.62 The N-terminal domain of hERG
plays a key role in the deactivation process.
Deleting amino acids 2 to 354 (2-354)63,64 or
2 to 373 (2-373)65 from the N-terminal domain
causes a ten-fold acceleration of hERG de-
activation. This effect is largely mediated by a
short stretch of amino acids (2–16) in the N-
terminal end of hERG.64,66 Studies by Robertson
and coworkers have provided important clues as
to how the N-terminal domain mediates the slow
deactivation process of the hERG channel.64,66
They show that the N-terminal peptide (amino
acids 2–16) can stabilize the hERG channel in
the open state, and an important part of the
docking site for the N-terminal domain is the
S4–S5 linker region of the activation gate.
A helix-loop-helix ‘‘Per-Arnt-Sim’’ (PAS) motif
is identified in the crystal structure of a peptide
corresponding to the first 135 amino acids of
the hERG channel.67 It is suggested that the
hydrophobic surface of the PAS motif can engage
in binding interaction with other regions of the
channel, and stabilize N-terminal domain binding
to the activation gate. LQT2-associated mutations
occurring around the PAS motif share a common
phenotype: acceleration of hERG deactiva-
tion.62 This is probably due to a disruption of
the PAS motif, thus destabilizing binding of the
N-terminal domain to the activation gate at
negative voltages.
Inactivation and recovery from inactivation
An important feature of the IKr/hERG channels is the
inward rectification property that limits outward
currents through the channels at positive voltages.
This reduces the amount of inward currents needed
to maintain the action potential’s plateau phase.
The inward rectification is due to a fast inactivation
process.63 hERG mutations in the pore-loop and in
the N-terminal half of the S6 domain can disrupt
the fast inactivation process.65,68–72 These are equi-
valent to the positions that are important for
Shaker’s C-type inactivation,68 suggesting a similar
mechanism underlying these two inactivation pro-
cesses. In the Shaker channel, the C-type in-
activation results from a concerted movement of
840 G.-N. Tseng
all four subunits,73,74 causing a rearrangement of
the outer mouth of the pore75 that either constricts
the pore and prevent ion fluxes, or alters the ion
selectivity in such a way that the channel cannot
conduct K currents under normal ionic conditions.76
Despite the similarity in structural elements in-
volved in the C-type inactivation between hERG
and Shaker, the uniquely fast kinetics of hERG
inactivation requires further consideration. It has
been proposed that the hERG’s outer mouth is
narrower and more flexible than that of the Shaker
channel, so that the outer mouth rearrangement
during hERG’s inactivation process requires a smal-
ler molecular motion, and is much faster, than that
during the Shaker’s C-type inactivation process.72,77
Clues for the difference in the outer mouth structure
between the hERG and Shaker channels can be
found by comparing the pore-loop sequences of
the two (Fig. 1). The Shaker sequence has double
tryptophans (WW) at the N-terminal end of the
pore-loop, and a tyrosine (Y) in the ‘‘signature
motif’’ (GYG) at the C-terminal end. According to
the crystal structure of KcsA, a model K channel,
hydrogen bonds can be formed in the 3-dimensional
structure around the outer mouth between the
nitrogens of ‘‘WW’’ and the hydroxyl groups of ‘‘Y’’
of the four subunits. These hydrogen bonds serve
as a layer of ‘‘molecular spring’’, that stretches
radially outward to hold the outer mouth open at
its proper diameter.54 In the hERG channel, the
‘‘WW’’ are replaced by ‘‘YF’’ (lacking nitrogens),
and the ‘‘Y’’ is replaced by ‘‘F’’ (lacking hydroxyl
groups). Therefore, the hydrogen bonds are missing
in hERG. This will weaken the ‘‘molecular spring’’,
resulting in a narrower but more flexible outer
mouth structure in the hERG channel.
The S5-P loop of hERG (39 amino acids) is longer
than those of most other Kv channels (5–10 amino
acids). Several LQT2-associated mutations oc-
curring here allow channel maturation and traf-
ficking to the cell surface, but cause defects in
channel function,5,78 Therefore the S5-P loop of
hERG may play an important role in channel func-
tion. Studies of mutations made in the center of
hERG’s S5-P loop suggest that this loop may interact
with the outer mouth of the pore, and participates
in the inactivation and ion selection processes.79
Ion permeation
As discussed above, the crystal structure of KcsA
suggests that in most Kv channels the hydrogen
bonds formed between tryptophan (W) nitrogens
at the N-terminal end of the pore-loop and the
hydroxyl groups of tyrosines (Y) in the ‘‘GYG’’
motif help maintain the outer mouth at its proper
dimension.54 Therefore, the selectivity filter of Kv
channels can better coordinate dehydrated K+ ions,
but not the smaller dehydrated Na+ ions. The high
affinity for K+ ion binding to the selectivity filter,
in conjunction with electrostatic repulsion between
two or three other K+ ions simultaneously residing
in the Kv channel pore, determines the channels’
high K+ selectivity against Na+ ions and yet main-
tains a high throughput rate of the pore. Such
hydrogen bonds are missing in the hERG channel
(Fig. 1). This may create a more flexible selectivity
filter in the hERG channel, making it prone to
lose its K+/Na+ selectivity. This may explain why
external Na+ ions can block the hERG pore with
an unusually high potency. The IC50 is 3 m (nom-
inally zero [K+]o),80 a potency much higher than
Na+ blockade of other Kv channels under com-
parable ionic conditions (IC50 in the range of tens
to 100 m).81
Subunit Composition of IKr Channels
It has been suggested that homomultimer hERG
channels expressed in heterologous systems have
a significantly slower deactivation rate than that of
native IKr channels in cardiac myocytes. Therefore,
other factors or subunits may be necessary to
recapitulate the gating properties of native IKr
channels. However, caution should be exercised
when comparing the hERG gating properties with
native IKr channels in cardiac myocytes. The
recording conditions (temperature,2 external pH,
45,82
concentrations of extracellular cations83,84)
can have important effects on the deactivation
rate of IKr/hERG channels. For the native IKr
channels, how the currents are isolated from
other interfering, and often much larger, currents
in cardiac myocytes may also influence the
apparent deactivation rate. Finally, under similar
recording conditions and using drug (E-4031 or
dofetilide)-sensitive currents as a measure of IKr,
there appear to be significant species variations
in the IKr deactivation rate (Table 1). Table 1
shows that the deactivation rate of homomultimer
hERG channels stably expressed in HEK 293 cells
is similar to that of IKr channels in human atrial
myocytes.2,30 However, it is not clear whether
HEK 293 cells express endogenous subunits that
can affect the deactivation rate of the exogenous
hERG channels.
 IKr: The hERG Channel
Role of splice variants of hERG subunits in IKr function?
N-terminal splice variants of hERG and mERG have
been identified.37,38 The ‘‘full-length’’ isoform (hERG
or mERG) has 396 amino acids in the N-terminus
and is designated as isoform 1a. The other isoform
(1b) has a much shorter (36 amino acids) and
divergent N-terminus. Isoform 1b lacks the
sequence of amino acids 2–16 and the PAS motif,
that are important for the slow deactivation process
of isoform 1a.64,66,67 Isoform 1b deactivates at a ten-
fold faster rate than that of isoform 1a.37 It has
been suggested that the two isoforms can form
heteromultimer channels.37 Therefore, if both iso-
forms are expressed in the same cell and can ran-
domly associate to form homo- and heteromultimer
channels, the resulting deactivation rate will depend
on the expression levels of the two isoforms. How-
ever, since the slow deactivation process of isoform
1a may require simultaneous binding of three or
four N-terminal domains to the activation gate,66
the phenotype will tend to be dominated by the
fast-deactivation rate.
It has been suggested that heteromultimer chan-
nels of isoforms 1a and 1b can better recapitulate the
gating kinetics of native IKr channels.37,38 Recently,
Nerbonne and colleagues used antibodies against the
N- and C-termini of hERG to probe the expression of
these two isoforms in adult human, rat and mouse
hearts.20 The anti-C antibody could recognize both
isoforms, while the anti-N antibody could recognize
1a but not 1b isoform. The results indicate that iso-
form 1a is expressed in adult hearts of all three spe-
cies, but there is no detectable expression of isoform
1b. These data raise questions about the significance
of isoform 1b in the IKr channel function in adult
hearts. However, isoform 1b may contribute to the
IKr channel function in neonatal hearts. For example,
a preliminary report shows that in ventricular myo-
cytes from 6–7 day old mice of homozygous mERG
1b knockout (−/−), the IKr current amplitude is
reduced and the IKr deactivation rate is dominated by
a slow rate.85 These data suggest that isoform 1b
makes a sizable contribution to the IKr channel func-
tion in these young animals. Developmental changes
in ERG isoform expression may also explain the find-
ing that in neonatal cat ventricular myocytes the IKr
deactivation rate is significantly faster than that in
adult heart.22
A C-terminal splice variant of hERG has been
identified in normal human heart.86 This subunit
(hERGUSO) does not form functional channels on
its own. However, when coexpressed with hERG,
hERGUSO suppresses the hERG current amplitude
and alters its gating kinetics (accelerating activation
841
and shifting the voltage-dependence by −8.8 mV).
The hERGUSO mRNA level is two-fold more abundant
than that of hERG. Therefore it has been suggested
that hERGUSO may play a role in determining the
current amplitude and gating kinetics of IKr in
cardiac myocytes. However, so far there has been
no biochemical evidence supporting the expression
of hERGUSO protein in the heart.
Role of regulatory
subunits in IKr/hERG channel
function?
The first indication that there may be regulatory
(
) subunits that can modulate the hERG channel
function came from a study on a mouse atrial cell
line (AT-1 cells).87 IKr is the major outward current
in AT-1 cells. Treating the cells with anti-sense
oligonucleotides against minK, and thus sup-
pressing the minK expression, reduced the IKr cur-
rent amplitude.87 The notion that minK can
augment the IKr current amplitude and modulate
its gating kinetics is further supported by the ob-
servation that in homozygous minK knockout (−/
−) mouse myocytes, the IKr amplitude is sig-
nificantly smaller and its deactivation rate is slower
than IKr in minK +/+ animals.88 Furthermore,
McDonald et al. showed that when coexpressed in
HEK 293 cells, hERG and minK can form a stable
complex, and the hERG current amplitude is aug-
mented relative to cells expressing hERG alone.89
However, so far there has been no biochemical
evidence for an association between minK and ERG
proteins in cardiac myocytes.
In 1999, Goldstein and co-workers identified three
gene families that encode minK-related peptides
(MiRPs). These MiRPs, along with minK, constitute
four KCNE families (KCNE1 to KCNE4, encoding
minK and MiRP1 to MiRP3).4 It is shown that in
vitro translated MiRP1 and hERG can form a stable
complex. MiRP1 exerts the following effects on the
hERG channel: accelerating deactivation, shifting
the voltage-dependence of activation in the positive
direction, reducing channel sensitivity to [K+]o
elevation, and sensitizing channel to IKr blockers (E-
4031 and clarithromycin).4 Furthermore, MiRP1
mutations have been identified that are linked to
congenital (LQT6) or acquired LQT syndrome.4,5,90
Therefore, it is suggested that MiRP1 is the
subunit
of native IKr channel in cardiac myocytes. However,
currently there is no biochemical evidence sup-
porting an association between native MiRP1 and
hERG proteins in cardiac myocytes.
Recently, MiRP2 has been shown to suppress
the expression of hERG in oocytes, suggesting yet
842 G.-N. Tseng
another
subunit that can potentially modulate
the hERG channel function.91 It is possible that
KCNE
subunits (minK, MiRP1 and MiRP2) can
engage in interactions with  subunits from more
than one gene family, and their role in native
Kv channel function may be determined by the
expression level and/or the subcellular distribution
of different Kv channel subunits. In support of this
notion, a recent preliminary report shows that
MiRP1 can physically associate with Kv4.2 (both
in vitro translated). When coexpressed in oocytes,
MiRP1 exerts profound effects on the gating kinetics
and pharmacology of Kv4.2.92
Structural Basis for the Unique
Pharmacological Properties of IKr/hERG
Channels
Methanesulfonanilides and other blockers
IKr/hERG channels are the target of highly specific
and potent class III anti-arrhythmic drugs, meth-
anesulfonanilides.7,93 The IKr/hERG channels can
also be blocked by a wide range of other therapeutic
agents with diverse chemical structures.94–101 These
unique pharmacological properties highlight the
importance of understanding the molecular de-
terminants of drug actions on the IKr/hERG chan-
nels.
Methanesulfonanilides block the hERG channel
from the intracellular side of the membrane102 and
binding primarily occurs in the open state.7,61 Un-
binding of some methanesulfonanilides (dofetilide,
MK-499) is very slow and incomplete at negative
voltages. These observations suggest that the clos-
ure of the activation gate at negative voltages not
only prevents methanesulfonanilides from gaining
access to their binding site, but also traps the drug
molecules once they are inside the channel pore.93
The ‘‘trapping’’ phenomenon is confirmed by a
recent study using a mutant hERG channel
(D540K) that can be opened by strong membrane
hyperpolarization59: MK-499 blockade of D540K
can be relieved at negative voltages.103
The elegant studies by Sanguinetti and coworkers
have revealed two important factors that contribute
to the unique pharmacological properties of IKr/
hERG channels.55,103 First, their data suggest that
the volume of hERG’s inner vestibule is larger than
those of most other Kv channels. Therefore, meth-
anesulfonanilides (e.g. MK-499, with a dimension
of 7×20Å) can be trapped within the inner ves-
tibule of the hERG channel without affecting the
deactivation kinetics.103 Structurally, the larger
inner vestibule can be explained by the lack of
proline residues in the S6 domain of the hERG
channel. In all other Kv channels, there are 1 or
2 proline residues in the carboxyl end of the S6
domains (Fig. 1). These proline residues cause a
sharp bend in the S6 helices,104 and may reduce
the volume of the inner vestibule. The lack of such
proline residues makes hERG’s S6 domain more
flexible, capable for forming a larger inner vestibule.
This favors drug binding to the hERG channel.
Second, the carboxyl-half of S6 in hERG has two
aromatic residues (Y652 and F656), that form part
of the ‘‘contact points’’ with inner mouth blockers
(Fig. 1).55,60 Homology modeling of the inner mouth
structure of hERG, based on the crystal structure
of KcsA, further suggests that the aromatic moieties
of methanesulfonanilides and other drugs (ter-
fenadine and cisapride) form electrostatic inter-
actions with these two aromatic residues by 
electron stacking.55 These two aromatic residues
are unique to hERG (the equivalent positions are
occupied by isoleucine or valine in other Kv chan-
nels, Fig. 1). They contribute to the channel’s high
affinity for drugs with aromatic moieties. Therefore,
the features of hERG’s S6 sequence play a central
role in determining the channel’s unique phar-
macological properties.
In addition to the S6 sequence, the fast in-
activation process of the hERG channel is also
important for the binding of certain (meth-
anesulfonanilides),60 but not all (vesnarinone,105
quinidine60), drug molecules. Many mutations that
disrupt the fast inactivation process of the hERG
channel also reduce channel sensitivity to meth-
anesulfonanilides.106,107 It is suggested that the fast
inactivation process of the hERG channel can in-
duce allosteric changes in the inner mouth region,
allowing the formation of a high affinity binding
site for methanesulfonanilides.60,108 Since a strong
depolarization (to +80 mV) reduces dofetilide bind-
ing relative to that at a modest depolarization
(+10 mV),61 it is further suggested that the high
affinity drug binding site is a ‘‘preinactivated’’
state.60
It has been shown that MiRP1 can enhance the
sensitivity of the hERG channel to E-4031 and
clarithromycin.4 MiRP1 also increases the degree
of tonic block and accelerates the development of
use-dependent block of hERG by E-4031.4 However,
MiRP1 does not affect the sensitivity of the hERG
channel to d-sotalol108 or dofetilide.109 The mech-
anism for the effects of MiRP1 on hERG phar-
macology, and the general implications of such
effects, are not clear at present.
 IKr: The hERG Channel
New concepts about drug actions on the IKr/hERG
channels
Several IKr/hERG blockers can increase the current
amplitude under certain conditions. First, E-4031
and some other IKr blockers can rescue defective
processes of protein trafficking that occur in many
LQT2-associated mutations.78,110 These drugs can
thus increase cell surface expression of these mutant
hERG proteins, and induce or increase the current.
Second, azimilide,101 almokalant,111 and nor-
propoxyphene100 can induce an ‘‘agonist’’ effect on
IKr/hERG under conditions of low drug con-
centrations and low depolarization voltages. At high
concentrations and stronger depolarizations, their
blocking effects dominate. The agonist effect of
azimilide on hERG has been analyzed in detail.101
Azimilide exerts this effect from the extracellular
side of the membrane. The binding site is away
from the outer mouth of the pore (probably in
the extracellular S3-S4 linker region). Azimilide’s
agonist effect is intimately related to the activation
process of the hERG channel. It is not clear whether
the other drugs share the same mechanism or a
similar binding site. These novel findings suggest the
possibility of designing chemicals that can enhance
hERG current amplitude (by correcting trafficking
defects or by inducing an ‘‘agonist’’ effect) without
the blocking effects. These agents may be useful for
congenital or acquired LQT syndrome.
Modulation of IKr/hERG Channel Function
by Changes in Cellular Milieu
Pathological conditions of the heart often cause
local extracellular K+ accumulation and acidosis.
Autonomic (e.g.
-adrenergic) activities may also
be elevated in an uneven fashion. These may lead
to heterogeneous changes in IKr current amplitude
and/or gating kinetics in different regions of the
heart. Long-term disease conditions of the heart
can also alter IKr expression that shows regional
variations. Heterogeneous changes in IKr function
under pathological conditions may contribute to
derangement of cardiac electrical activity and
arrhythmogenesis.
Acute IKr/hERG modulation by changes in the
extracellular cation composition
Extracellular K+ accumulation
Elevating [K+]o paradoxically increases outward cur-
rents through the IKr/hERG channels despite a de-
843
crease in the driving force for outward currents.3,42
This occurs through a combination of several mech-
anisms. First, elevating [K+]o relieves IKr/hERG chan-
nels from the inactivated state by hindering the
conformational changes in the outer mouth region
necessary for the inactivation process.43,112–114 Since
the inactivation process is the primary limiting factor
for outward IKr/hERG currents at depolarized volt-
ages, this is the most important mechanism by which
elevating [K+]o increases the outward IKr/hERG cur-
rent amplitudes. Second, elevating [K+]o can relieve
channel blockade by extracellular Na+ ions.80 Third,
elevating [K+]o increases the single channel con-
ductance of IKr/hERG channels.102,115–117 The latter
two mechanisms will lead to an increase of both out-
ward and inward currents through the IKr/hERG
channels.
Extracellular acidosis
Elevating [H+]o (extracellular acidification from
pH 7.4 to 6.4 or 6) induces a marked acceleration
of IKr/hERG deactivation.45,82,118,119 This occurs with
little or no changes in the current amplitude or
the voltage-dependence of other gating processes,
indicating that the underlying mechanism is not
pore blockade or screening of negative surface
charges by protons. The effect of elevating [H+]o on
hERG deactivation is reduced when the cytoplasmic
N-terminal domain (amino acids 2–354) is re-
moved.82 This effect can also be prevented by pre-
teating the hERG channel with extracellular
diethylpyrocarbonate (DEPC), that can covalently
modify the side chains of histidine and cysteine.120
These observations suggest that protonation of res-
idues on the hERG channel surface can induce
allosteric changes in the channel, destabilizing bind-
ing of the N-terminal domain to the activation
gate at negative voltages and accelerating channel
deactivation.
Acute modulation of IKr/hERG channel function by

-adrenergic stimulation
The amino acid sequence of hERG shows that there
are four putative cAMP-dependent protein kinase
(PKA) phosphorylation sites and a cyclic nucleotide
binding domain (NBD) in the cytoplasmic regions
of the channel. These features suggest that the
hERG channel may be modulated by PKA phos-
phorylation and probably by direct cAMP binding.
Indeed, studies on the hERG channel expressed in
oocytes46,121 and in mammalian cells47 show that
844
PKA activation induces a positive shift in the volt-
age-dependence of hERG activation, an acceleration
of deactivation, and a decrease in the current amp-
litude. Removing all four PKA phosphorylation sites
prevents such effects.46,47 Interestingly, in mam-
malian cells expressing the hERG mutant with all
four PKA phosphorylation sites removed, elevating
the intracellular cAMP level induces a negative shift
in the voltage-dependence of channel activation,
i.e. opposite to the voltage shift caused by PKA
phosphorylation.47 Coexpression of minK or MiRP1
accentuates this cAMP-induced voltage shift.47 Bio-
chemical data further support the notion that cAMP
can directly bind to the hERG channel protein.47
These data suggest that the hERG channel can be
modulated by
-adrenergic stimulation in a complex
manner. Previously, it was shown that the native
IKr channel in guinea pig ventricular myocytes could
not be modulated by
-adrenergic stimulation.122
This could be due to species variations in the ERG
amino acid sequence. The important issue of
whether and how native IKr channels in human
cardiac myocytes can be modulated by
-adrenergic
stimulation requires further investigation.
Long-term modulation of IKr/hERG function by disease
conditions of the heart
Disease conditions of the heart can affect the func-
tion and/or expression of ion channels.123,124 This
information is important for the understanding of
pathophysiology and arrhythmogenesis of the dis-
eased hearts, and for the design of therapeutic
strategies for these conditions. Currently, relatively
little is known about how the IKr channel function
and the expression of its subunits are modulated
by chronic disease of the heart. The following is a
brief summary of what is available in the literature.
In canine epicardial myocytes isolated from the
infarct zone 5 days after a total occlusion of the left
anterior descending coronary (LAD), the IKr current
density is reduced and the activation kinetics appears
to be accelerated.49 The ERG mRNA level in infarct
zone tissue is reduced on day 2 and day 5 post-in-
farction, suggesting that the reduced IKr current
density is due to a decrease in ERG protein expression.
However, it is not clear whether there are changes
in other (regulatory) subunits that can account for
the altered IKr gating kinetics. The ERG mRNA level
is not altered in noninfarct zone tissue on either day 2
or day 5, suggesting regional variations in IKr current
density in these infarcted canine hearts. As discussed
above, canine subendocardial Purkinje myocytes ex-
press a unique E-4031-sensitive outward current
G.-N. Tseng
that has gating kinetics similar to that of EAG, and
may represent a novel ERG isoform.39 The current
density of this E-4031 sensitive current is increased
in subendocardial Purkinje myocytes in infarct zone
48 h after LAD occlusion.39 The upregulation of an
E-4031-sensitive current in Purkinje myocytes may
increase the risk of pro-arrhythmic effects of class III
anti-arrhythmic drugs in the setting of myocardial
infarction. In a canine model of biventricular hyper-
trophy induced by chronic complete atrioventricular
block, the IKr current density is reduced in myocytes
from the right ventricle.48 However, IKr is not affected
in myocytes from the left ventricle,48 indicating
regional variations in IKr changes in the hyper-
trophied canine heart. In myocytes from hyper-
trophied right125 and left126 ventricles of the cat, the
density of the delayed rectifier current (IK) is reduced.
This is accompanied by a slowing of IK activation and
an acceleration of deactivation. The identity of IK in
these studies is not clear. However, IKr may be the
major component.21 The IKr channel function can
also be altered after long-term drug therapy. For ex-
ample, in guinea pig ventricular myocytes after
chronic amiodarone therapy (80 mg/kg ip daily for
7 days), the IKr current density (along with those of
IKs and IK1) is severely suppressed.127
Questions for Future Studies
The heterogeneity in IKr channel function plays an
important role in maintaining the cardiac electrical
stability under normal conditions. Disturbances in
IKr channel function, either due to genetic defects
or due to drug effects, increase the risk of life
threatening arrhythmias. The intense research ef-
forts on the IKr channel during the last decade,
especially after the identification of the hERG gene,
have provided tremendous insights into the role of
this channel in action potential repolarization, its
structure–function relationship and the mech-
anisms of its unique pharmacological properties.
However, there remain important questions that
require further investigation: (1) the role of splice
variants of hERG and the role of KCNE
subunits
in native IKr function in cardiac myocytes need to
be defined; (2) the effects of chronic pathological
conditions of the heart (e.g. hypertrophy, infarction,
and heart failure) on the function and expression
of IKr channels need to be characterized, and the
underlying mechanisms need to be identified; (3)
the structure–function relationship of the hERG
channel, alone or with coassembled
subunits,
needs further analysis. In particular, the 3-di-
mensional structure of the hERG pore can be better
 IKr: The hERG Channel
defined now, using the approach of homology mod-
eling based on the crystal structure of the KscA pore.
However, this endeavor should take into careful
consideration the unique features of the pore-loop
sequence, the S6 sequence, and the role of the
S5-P loop in the hERG channel, as suggested by
mutagenesis studies. Finally, agents that can in-
crease the IKr current amplitude may be useful for
the congenital or acquired long QT syndrome. It
is expected that the better defined 3-dimensional
structure of the hERG pore will facilitate the design
of new therapeutic agents that can increase the
current amplitude (by correcting trafficking defects
of mutant hERG proteins or by inducing an agonist
effect) without pore blocking effects.
References
1. H JC, L AJ, W HJ. Time course
and voltage dependence of expressed HERG current
compared with native ‘‘rapid’’ delayed rectifier K
current during the cardiac ventricular action po-
tential. Pflugers Arch 1998; 436: 843–853.
2. Z J, G Q, Y B, F Z, M JC, R-
 GA, J CT. Properties of HERG chan-
nels stably expressed in HEK 293 cells studied
at physiological temperature. Biophys J 1998; 74:
230–241.
3. S MC, J C, C ME, K
MT. A mechanistic link between an inherited and
an acquired cardiac arrhythmia: HERG encodes the
IKr potassium channel. Cell 1995; 81: 299–307.
4. A GW, S F, S I, B ME, L
MH, T KW, K MT, G SAN.
MiRP1 forms IKr potassium channels with HERG
and is associated with cardiac arrhythmia. Cell
1999; 97: 175–187.
5. S I, S J, T KW, L MH,
P SG, R JL, M AJ, S PJ,
T JA, V GM, K MT. Spectrum of
mutations in long-QT syndrome genes KvLQT1,
HERG, SCN5A, KCNE1, and KCNE2. Circulation
2000; 102: 1178–1185.
6. S MC, J NK. Two components
of cardiac delayed rectifier K+ current. Differential
sensitivity to block by class III anti-arrhythmic
agents. J Gen Physiol 1990; 96: 195–215.
7. S PS, C ME, K MT, S
MC. Class III anti-arrhythmic drugs blocks HERG,
a human cardiac delayed rectifier K+ channel.
Open-channel block by methanesulfonanilides. Cir
Res 1996; 78: 499–503.
8. N JM. Molecular basis of functional voltage-
gated K+ channel diversity in the mammalian myo-
cardium. J Physiol 2000; 525: 285–298.
9. M JS, S MC. Biophysical prop-
erties and molecular basis of cardiac rapid and slow
delayed rectifier potassium channels. Cell Physiol
Biochem 1999; 9: 201–216.
10. R DM, B JR. A plethora of mechanisms
in the HERG-related long QT syndrome. Genetics
845
meets electrophysiology. Cardiov Res 1999; 44:
242–246.
11. S MC. Dysfunction of delayed rectifier
potassium channels in an inherited cardiac arrhyth-
mia. Annal New York Academy of Sciences 1999;
868: 406–413.
12. W HJ, H JC. Familial and acquired long
QT syndrome and the cardiac rapid delayed rectifier
potassium current. Clin Exp Pharm Physiol 2000;
27: 753–766.
13. P SG, B J, H RNW, H
W, J HJ, K AG, MK WJ, R
DM, R Y, S K, S PJ, T
JA, W A. Genetic and molecular basis of cardiac
arrhythmias. Impact on clinical management. Eur
Heart J 1999; 20: 174–195.
14. S MC, J NK. Delayed rectifier
outward K+ current is composed of two currents
in guinea pig atrial cells. Am J Physiol 1991; 260:
H393–H399.
15. L S, R RL, C DL, W S,
S HC. Activation and inactivation kinetics of
an E-4031-sensitive current from single ferret atrial
myocytes. Biophys J 1996; 70: 2704–2715.
16. D MP, A RH, D P, K S,
C KR, K RS. Developmental changes in ionic
channel activity in the embryonic murine heart.
Cir Res 1996; 78: 15–25.
17. W L, F Z-P, K CS, S RS, D
HJ. Developmental changes in the delayed rectifier
K+ channels in mouse heart. Circ Res 1996; 79:
79–85.
18. B P, A R, N B, S C-M,
B TR, J B, A TM, K J, D-
G LJ, S W, C T. Inhibition of
cardiac delayed rectifier K+ current by over-
expression of the long-QT syndrome HERG G628S
mutation in transgenic mice. Circ Res 1998; 83:
668–678.
19. W RS, G GA, W RT, D JE,
MK D, C IS. Tissue and species dis-
tribution of mRNA for the IKr-like K+ channel, erg.
Circ Res 1997; 80: 261–268.
20. P AL, S BK, B AT, P K,
 W DR, S A, N JM. Ex-
pression of distinct ERG proteins in rat, mouse, and
human heart. Relation to functional IKr channels.
J Biol Chem 2000; 275: 5997–6006.
21. F CH, C TJ. Block of delayed rectifier
potassium current, IK, by flecainide and E-4031
in cat ventricular myocytes. Circulation 1990; 82:
289–293.
22. B-M H, E A, S-
C JA. Developmental differences in delayed
rectifying outward current in feline ventricular
myocytes. Am J Physiol 2000; 278: H484–H492.
23. S JJ, J NK, J B, F K,
G PJ, R B, S R, F B. IK
of rabbit ventricle is composed of two currents:
evidence for IKs. Am J Physiol 1996; 271: H2477–
H2489.
24. M JS, H JC. An investigation of the
role played by the E-4031-sensitive (rapid delayed
rectifier) potassium current in isolated rabbit atrio-
ventricular nodal and ventricular myocytes. Pflu-
gers Arch 2000; 438: 843–850.
846 G.-N. Tseng
25. C JM, S KW, G WR. Repolarizing
K+ currents in rabbit heart Purkinje cells. J Physiol
1998; 508: 811–823.
26. G GA. Two components of delayed rectifier
current in canine atrium and ventricle. Does IKs
play a role in the reverse rate dependence of class
III agents? Circ Res 1996; 78: 26–37.
27. L D-W, A C. Characteristics of the
delayed rectifier current (IKr and IKs) in canine vent-
ricular epicardial, midmyocardial, and endocardial
myocytes. A weaker IKs contributes to the longer
action potential of the M cell. Circ Res 1995; 76:
351–365.
28. F J, Y L, W Z, N S. Ionic mechanisms
of regional action potential heterogeneity in the
canine right atrium. Circ Res 1998; 83: 541–551.
29. G GA. Characterization and functional con-
sequences of delayed rectifier current transient in
ventricular repolarization. Am J Physiol 2000; 278:
H806–H817.
30. W Z, F B, N S. Rapid and slow
components of delayed rectifier current in human
atrial myocytes. Cardiov Res 1994; 28: 1540–1546.
31. L G-R, F J, Y L, C M, N S. Evi-
dence for two components of delayed rectifier K+
current in human ventricular myocytes. Circ Res
1996; 78: 689–696.
32. B MV, M MJ, R KA,
S HC. Regional localization of ERG, the chan-
nel protein responsible for the rapid component of
the delayed rectifier, K+ current in the ferret heart.
Circ Res 1997; 81: 128–135.
33. C J, K K, L W, T Y, T J,
K I. Heterogeneous distribution of the two
components of delayed rectifier K+ current: a po-
tential mechanism of the pro-arrhythmic effects of
methanesulfonanilide class III agents. Cardiov Res
1999; 43: 135–147.
34. M MC, B SM, H G. Regional differences
in action potential characteristics and membrane
currents of guinea-pig left ventricular myocytes.
Exp Physiol 1998; 83: 747–761.
35. F T, K S, F N, B AL,
M RJ. Potassium rectifier currents differ in
myocytes of endocardial and epicardial origin. Circ
Res 1992; 70: 91–103.
36. C JR, O A, P T, S BI,
S A. A quantitative description of the E-4031-
sensitive repolarization current in rabbit ventricular
myocytes. Biophys J 1995; 69: 1830–1837.
37. L B, T MC, N KP, B AK,
C NG, G DJ, J NA, S CA,
R GA. Two isoforms of the mouse ether-
a-go-go-related gene coassemble to form channels
with properties similar to the rapidly activating
component of the cardiac delayed rectifier K+ cur-
rent. Circ Res 1997; 81: 870–878.
38. L-M JP, K C, W L, D HJ. Elec-
trophysiological characterization of an alternatively
processed ERG K+ channel in mouse and human
hearts. Circ Res 1997; 81: 719–726.
39. P JMB, B PA. Reduced inward rectifying
and increased E-4031-sensitive K+ current density
in arrhythmogenic subendocardial Purkinje myo-
cytes from the infarcted heart. J Cardiov Electro-
physiol 1998; 9: 299–311.
40. S WR, T C-Y, M AF, H K-B,
P DM. Mg2+ modulates voltage-dependent
activation in Ether-a-go-go potassium channels by
binding between transmembrane segments S2 and
S3. J Gen Physiol 2000; 116: 663–677.
41. J NK, S MC. Rate-dependent
prolongation of cardiac action potentials by a meth-
anesulfonilide class III anti-arrhythmic agent. Spe-
cific block of rapidly activating delayed rectifier K+
current by dofetilide. Circ Res 1993; 72: 75–83.
42. S MC, J NK. Role of external
Ca2+ and K+ in gating of cardiac delayed rectifier
K+ currents. Pflugers Archiv 1992; 420: 180–186.
43. Y T, S DJ, R DM. Rapid inactivation
determines the rectification and [K+]o dependence
of the rapid component of the delayed rectifier
K+ current in cardiac cells. Circ Res 1997; 80:
782–789.
44. S F, C E. Delayed K+ current and
external K+ in single cardiac Purkinje cells. Am J
Physiol 1989; 257: C1086–C1092.
45. V J, C E. The effect of external pH
on the delayed rectifying K+ current in cardiac
ventricular myocytes. Pflugers Arch 2000; 439:
739–751.
46. T D, Z W, K CA, K S,
S W, K W, K J. Deletion of protein
kinase A phosphorylation sites in the HERG potas-
sium channel inhibits activation shift by protein
kinase A. J Biol Chem 1999; 274: 27457–27462.
47. C J, M Y, P E, F GI, MD
TV. Cyclic AMP regulates the HERG K+ channel by
dual pathways. Cur Biol 2000; 10: 671–674.
48. V PGA, S KR, V MA, S
RLHMG, L JDM, C E, W
HJJ. Downregulation of delayed rectifier K+ currents
in dogs with chronic complete atrioventricular
block and acquired Torsades de Pointes. Circulation
1999; 100: 2455–2461.
49. J M, C C, Y J-A, B PA, T G-N.
Delayed rectifier K currents have reduced amp-
litudes and altered kinetics in myocytes from in-
farcted canine ventricle. Cardiov Res 2000; 48:
34–43.
50. R DM, G AL. Structure and function
of cardiac sodium and potassium channels. Am J
Physiol 1997; 273: H511–H525.
51. G HR, D SR. Structural models of Na+,
Ca2+ and K+ channels. Soc Gen Physiol series 1995;
50: 1–16.
52. K M, L P, L R. Molecular
determinants of ion conduction and inactivation in
K+ channels. Am J Physiol 1995; 268: C535–C556.
53. H L, L Z, A T, MK R.
Mutations in the K+ channel signature sequence.
Biophys J 1994; 66: 1061–1067.
54. D DA, C JM, P RA, K A,
G JM, C SL, C BT, MK R. The
structure of the potassium channel: molecular basis
of K+ conduction and selectivity. Science 1998; 280:
69–77.
55. M JS, C J, L M, C C, S-
 MC. A structural basis for drug-induced
long QT-syndrome. Proc Natl Acad Sci 2000; 97:
12329–12333.
56. H M, S KS, Y G. The activation
 IKr: The hERG Channel
gate of a voltage-gated K+ channel can be trapped
in the open state by an intersubunit metal bridge.
Neuron 1998; 21: 617–621.
57. S C-C, K KG, K GE. Role of trans-
membrane segment S5 on gating of voltage-de-
pendent K+ channels. J Gen Physiol 1997; 109:
767–778.
58. Z Y-Y, J M, L S, T G-N. Stabilizing
channel’s open state by a hydrophobic residue in
the S6 domain of rKv1.4. Pflugers Archiv 1998;
437: 114–122.
59. S MC, X QP. Mutations of the S4–S5
linker alter activation properties of HERG potassium
channels expressed in Xenopus oocytes. J Physiol
1999; 514: 667–675.
60. L-M JP, D Y, T GQ, D HJ. Mo-
lecular determinant of high-affinity dofetilide bind-
ing to HERG1 expressed in Xenopus oocytes:
involvement of S6 sites. Mol Pharm 2000; 57:
367–374.
61. S DJ, C A. High affinity open chan-
nel block by dofetilide of HERG expressed in a
human cell line. Mol Pharm 1996; 49: 949–955.
62. C J, Z A, S I, K MT, S-
 MC. Long QT syndrome-associated mut-
ations in the Per-Arnt-Sim (PAS) domain of HERG
potassium channels accelerate channel de-
activation. J Biol Chemi 1999; 274: 10113–10118.
63. S PS, C ME, Z A, K MT, San-
guinetti MC. Fast inactivation causes rectification of
the IKr channel. J Gen Physiol 1996; 107: 611–619.
64. W J, T MC, Z AM, R GA.
Regulation of deactivation by an amino terminal
domain in Human ether-a-go-go gene potassium
channels. J Gen Physiol 1998; 112: 637–647.
65. S R, H SH. Molecular de-
terminants for activation and inactivation of HERG,
a human inward rectifier potassium channel. J
Physiol 1996; 493: 635–642.
66. W J, M CD, R GA. Dynamic control
of deactivation gating by a soluble amino-terminal
domain in HERG K+ channels. J Gen Physiol 2000;
115: 749–758.
67. M C JH, L A, C SL, C BT, L
M, MK R. Crystal structure and functional
analysis of the HERG potassium channel N-ter-
minus: an eukaryotic PAS domain. Cell 1998; 95:
649–655.
68. H T, Z WN, A RW. Two types
of inactivation on Shaker K+ channels: effects of
alterations in the carboxy-terminal region. Neuron
1991; 7: 547–556.
69. S PL, B T, Y G. The inward
rectification mechanism of the HERG cardiac potas-
sium channel. Nature 1996; 379: 833–836.
70. H IM, T MC, R GA. Transfer
of rapid inactivation and sensitivity to the class III
anti-arrhythmic drug E-4031 from HERG to M-eag
channels. J Physiol 1998; 511: 3–14.
71. Z A, X QP, S MC. A mutation in the
pore region of HERG K+ channels expressed in
Xenopus oocytes reduces rectification by shifting the
voltage dependence of inactivation. J Physiol 1998;
509: 129–137.
72. F J-S, J M, D W, MD TV, T
G-N. Effects of outer mouth mutations on hERG
847
channel function: a comparison with similar mut-
ations in Shaker. Biophys J 1999; 76: 3128–3140.
73. P G, S Z-F, T L-W, D C. C-type
inactivation of a voltage-gated K+ channel occurs
by a cooperative mechanism. Biophys J 1995; 69:
896–903.
74. O EM, Z WN, H T, H
SH, H J, A RW. Cooperative subunit inter-
actions in C-type inactivation of K channels. Biophys
J 1995; 69: 2449–2457.
75. Y G, S D, C T-Y, J ME. An
engineered cysteine in the external mouth of a K+
channel allows inactivation to be modulated by
metal binding. Biophys J 1994; 66: 1068–1075.
76. S JG, K L, R MD, H
SH. Ion conduction through C-type inactivated
Shaker channels. J Gen Physiol 1997; 110: 539–
550.
77. D W, J M, Z S, T G-N. Differences in
the effects and state-dependence of MTS modi-
fication of an outer mouth cysteine residue reveal
differences between HERG and Shaker in structural
rearrangements involved in C-type inactivation.
Circulation 1999; 100: I–842.
78. Z Z, G Q, E ML, J CT. HERG
channel dysfunction in human long QT syndrome.
J Biol Chem 1998; 273: 21061–21066.
79. D W, J M, T G-N. Allosteric effects of
mutations in the extracellular S5-P loop on the
gating and ion permeation properties of hERG.
Pflugers Archiv 1999; 439: 141–149.
80. N H, J JPJ, P CI, B
JR. A sensitive mechanism for cation modulation
of potassium current. Nature Neuroscience 2000; 3:
429–430.
81. K L, I D, LT J, K SJ. The interaction
of Na+ and K+ in voltage-gated potassium channels.
Evidence for cation binding sites of different affinity.
J Gen Physiol 1998; 111: 195–206.
82. J M, D W, T G-N. Mechanism for the
effects of extracellular acidification on hERG chan-
nel function. Am J Physiol 1999; 277: H1283–
H1292.
83. J JP, M FM, B PB. Human ether-
a-go-go-related gene K+ channel gating probed with
extracellular Ca2+. Evidence for two distinct voltage
sensors. J Gen Physiol 1999; 113: 565–580.
84. H W-K, K I, L CO, Y JB, L SH, E
YE. Blockade of HERG channels expressed in Xeno-
pus laevis oocytes by external divalent cations. Bio-
phys J 1999; 76: 1959–1971.
85. L-M JP, S SL, S KJ, T K,
R DE, D HJ. Selective knock-out of the
ERG1B potassium channel: a mouse model for the
long-QT syndrome. Circulation 2000; 102: II–285.
86. K S, S DJ, R A, R DM. A
K+ channel splice variant common in human heart
lacks a C-terminal domain required for expression
of rapidly activating delayed rectifier current. J Biol
Chem 1998; 273: 27231–27235.
87. Y T, K S, R DM. Anti-minK
antisense decreases the amplitude of the rapidly
activating cardiac delayed rectifier K+ current. Circ
Res 1995; 77: 1246–1253.
88. K S, Y T, A ME, W A,
848 G.-N. Tseng
N KD, M MA, R DM. Re-
placement by homologous recombination of the
minK gene with lacZ reveals restriction of minK
expression to the mouse cardiac conduction system.
Circ Res 1999; 84: 146–152.
89. MD TV, Y Z, M Z, P E, M
MB, W K-W, G SAN, F GI. A
minK–HERG complex regulates the cardiac potas-
sium current IKr. Nature 1997; 388: 289–292.
90. S F, A GW, W J, M KT, S
S, S PJ, P SG, R DM, G AL,
G SAN. A common polymorphism as-
sociated with antibiotic-induced cardiac arrhyth-
mia. Proc Natl Acad Sci 2000; 97: 10613–10618.
91. S BC, W S, F S, B M,
W R, G R, J TJ. A constitutively
open potassium channel formed by KCNQ1 and
KCNE3. Nature 2000; 403: 196–199.
92. T G-N, A GW, J M. hMiRP1 mod-
ulates the gating kinetics and pharmacology of
transient outward current. Circulation 2000; 102:
II–285.
93. C E. Voltage- and time-dependent block of
the delayed K+ current in cardiac myocytes by
dofetilide. J Pharm Exp Ther 1992; 262: 809–817.
94. S H, W S, B AE. Blockade
of HERG channels expressed in Xenopus oocytes by
the histamine receptor antagonists terfenadine and
astemizole. FEBS Letters 1996; 385: 77–80.
95. R ML, D R, B AM. HERG, a primary
human ventricular target of the nonsedating anti-
histamine terfenadine. Circulation 1996; 94: 817–
823.
96. Z S, Z Z, G Q, M JC, J
CT. Mechanism of block and identification of the
verapamil binding domain to HERG potassium
channels. Circ Res 1999; 84: 989–998.
97. M S, Z Z, G Q, J CT. Block-
ade of the HERG human cardiac K+ channel by
the gastrointestinal prokinetic agent cisapride. Am
J Physiol 1997; 273: H2534–H2538.
98. S H, S R, H SH, A-
 B, L F, B AE. The inhibitory effect
of the antipsychotic drug haloperidol on HERG
potassium channels expressed in Xenopus oocytes.
Br J Pharm 1997; 120: 968–974.
99. W H-Z, S H, L S-J, W Z. Inactivation
gating determines nicotine blockade of human
HERG channels. Am J Physiol 1999; 277: H1081–
H1088.
100. U C, D P, T J. Norpropoxyphene-
induced cardiotoxicity is associated with changes
in ion-selectivity and gating of HERG currents.
Cardiov Res 1999; 44: 568–578.
101. J M, D W, F J-S, T G-N. Use-dependent
‘agonist’ effect of azimilide on the hERG channel. J
Pharm Exp Ther 1999; 291: 1324–1336.
102. K J, L AE, W B, B AM. Mo-
lecular physiology and pharmacology of HERG.
Single-channel currents and block by dofetilide.
Circulation 1996; 94: 2572–2579.
103. M JS, C J, S MC. Trapping
of a methanesulfonanilide by closure of the HERG
potassium channel activation gate. J Gen Physiol
2000; 115: 229–239.
104.  C D, H M, L Y, Y G.
Blocker protection in the pore of a voltage-gated
K+ channel and its structural implications. Nature
2000; 403: 321–325.
105. K K, S MC, T Y, H M, K-
 I. Vesnarinone selectively blocks open HERG
channels and block is not influenced by in-
activation. Circulation 2000; 102: II–355.
106. W S, M MJ, L S, S HC, R-
 RL. Modulation of HERG affinity for E-4031 by
[K+]o and C-type inactivation. FEBS Letters 1997;
417: 43–47.
107. F E, J W, K J, B A,
B AM. Molecular determinants of dofetilide
block of HERG K+ channels. Circ Res 1998; 82:
386–395.
108. N H, M FM, J JPJ, J DC,
P SS, Y ICH, T GF, B JR. Probing
the interaction between inactivation gating and d-
sotalol block of HERG. 2000; Circ Res 87: 1012–
1018.
109. W M, H T, N S. hMiRP1-HERG
association does not account for discrepancies with
native IKr channels in response to class III drugs.
Circulation 2000; 102: II–24.
110. Z Z, G Q, J CT. Correction of defective
protein trafficking of a mutant HERG potassium
channel in human long QT syndrome. Phar-
macological and temperature effects. J Biol Chem
1999; 274: 31123–31126.
111. C E. Use-dependent block and use-de-
pendent unblock of the delayed rectifier K+ current
by almokalant in rabbit ventricular myocytes. Circ
Res 1993; 73: 857–868.
112. B T, Y G. Modulation of K+ current
by frequency and external [K+]: A tale of two
inactivation mechanisms. Neuron 1995; 15: 951–
960.
113. B T, Y G. Use-dependent blockers
and exit rate of the last ion from the multi-ion pore
of a K+ channel. Science 1996; 271: 653–656.
114. W S, L S, M MJ, S HC, R-
 RL. A quantitative analysis of the activation
and inactivation kinetics of HERG expressed in
Xenopus oocytes. J Physiol 1997; 502: 45–60.
115. S T. Conductance and kinetics of delayed
rectifier potassium channels in nodal cells of the
rabbit heart. J Physiol 1987; 387: 227–250.
116. Z A, C ME, K MT, S MC.
Single HERG delayed rectifier K+ channels ex-
pressed in Xenopus oocytes. Am J Physiol 1997;
272: H1309–H1314.
117. V MW,  G ACG, B LN.
Single delayed rectifier channels in the membrane
of rabbit ventricular myocytes. Circ Res 1993; 72:
865–878.
118. B J, C M, D P. Modulation of
HERG potassium channel properties by external
pH. Pflugers Arch 1999; 438: 419–422.
119. A JMB, H J, D M, T SM,
J J. Proton and zinc effects on HERG currents.
Biophys J 1999; 77: 282–298.
120. M EW. Modification of histidyl residues in pro-
teins by diethylpyrocarbonate. Meth Enzymology
1977; 47: 431–443.
121. K J, K C, T D, Y X, B
J, K W. HERG potassium channel activation
 IKr: The hERG Channel
is shifted by phorbol esters via protein kinase A-
dependent pathways. J Biol Chem 1998; 273:
25285–25291.
122. S MC, J NK, S A, S
PKS. Isoproterenol antagonizes prolongation of re-
fractory period by the class III anti-arrhythmic
agent E4031 in guinea pig myocytes. Mechanism
of action. Circ Res 1991; 68: 77–84.
123. T GF, M E. Electrophysiological re-
modeling in hypertrophy and heart failure. Cardiov
Res 1999; 42: 270–283.
124. P JMB, B PA. Electrical remodeling in
ischemia and infarction. Cardiov Res 1999; 42:
284–297.
849
125. K RB, H SR. Outward currents in nor-
mal and hypertrophied feline ventricular myocytes.
Am J Physiol 1989; 256: H1450–H1461.
126. F T, M RJ, F N, K
S, B AL. Metabolic inhibition of ICa,L and IK
differs in feline left ventricular hypertrophy. Am J
Physiol 1994; 266: H1121–H1131.
127. B RF, L G-R, G R, N S. Electro-
physiologic effects of chronic amiodarone therapy
and hypothyroidism, alone and in combination, on
guinea pig ventricular myocytes. J Phar Exp Ther
1999; 289: 156–165.