Cellular and Molecular Biology

Phosphatidylinositol
Biphosphate
Pathway
Pathway
Ghimire, Priti
Salumbre, Renz
Surquia, Joseph
 Inositol Phospholipids
• Discovered by Hokin et
al in the 50s

Also known as
phosphatidylinositol (PI)

• First hypothesized as a
key player in hormone
action

Accepted as a second
messenger after 30
years of study
Inositol Phospholipids

It is the substrate for a

large number of
enzymes which are
involved in cell
signaling because it can
be phosphorylated by a
variety of kinases on
the hydroxyl groups 3, 4
and 5 on the inositol
ring in seven different
combinations.
Different IPs involved in
Cell Signaling
Cell Signaling
 Phosphatidyl Inositol
Biphosphate
• PIP2
• Located at the inner half of the lipid bilayer
• A precursor in the production of inositol trisphosphate (IP3 or
InsP3) and

diacylglycerol (DAG)
• It is hydrolyzed by phospholipase C (PLC) to produce such
secondary messengers

Only activated when the stimulatory G protein responded a

certain stimulatory signal.
Typical PIP2 Pathway
 The Role of Ca as
++

Intracellular Messenger
 Calcium Ions
• Significant Role in various cellular activities

muscle contraction
• cell division
• secretion

fertilization
• synaptic transmission
• metabolism

transcription
• cell movement

cell death
 The Role of Ca as ++

Intracellular Messenger
Extracellular
Message

Cell
Surface

high Ca++
Concentration
 The Role of Ca as ++

Intracellular Messenger

• Calcium ion concentration regulated by the activity of

calcium pumps and ion channels located within the
membranes that surround the compartment

Calcium concentration of a resting cell is maintained at low

levels
• Whereas, within the lumen or in the extracellular space,
calcium concentration is higher than that of the cytosol

Abnormal elevation of calcium ions can occur in brain cells

and lead to stroke and eventually death
IP3 and Voltage-Gated Ca ++

Channels
Extracellular messenger molecule +
GPCR

Activation of Phospholipase C-β

phosphoinositide PIP2 is split

IP3 molecule released

Calcium channels in the ER open

increase in cytosolic [Ca++]

 IP3 and Voltage-Gated Ca ++

Channels

• RTK activate members of the phospholipase C-ϒ subfamily

possess SH2 domain that allows them to bind to

activated and phosphorylated RTK
• 4 PLC -carry out the same reaction producing IP3 and
linking cell surface receptors to an increase in
cytoplasmic calcium ions

PLC-ϒ -activated by calcium ions

• PLC-ε -activated by Ras-GTP
IP3 and Voltage-Gated Ca ++

Channels
nerve impulse

depolarization of plasma membrane

opening of voltage-gated channels

influx of Ca++ ions

 Visualizing Ca ++

• Understanding the role of Ca++ ions was acheived through

the development of indicator molecules that emit light in the
presence of free calcium

Fluorescence microscopy and computerized-imaging

techniques
• Calcium-mediated responses are readily identifiable
 Types of Ca ion channels in
++

the ER membrane
• Two main types
• IP3 receptors
• Ryanodine receptors (RyRs)
• bind toxic plant alkaloid ryanodine
 Ryanodine Receptors

• Found in excitable cells

• In skeletal and cardiac muscle cells, they mediate the rise in
Ca++ levels after an arrival of APs
• Can be opened by many agents
• Calcium-Induced Calcium Release (CICR)
 Calcium Wave Induced by
Fertilizing Sperm
• Propagated wave of calcium release spreads through the
entire cytoplasmic compartment

Induced by the sperms contact with the egg’s plasma

membrane
• Sudden rise in calcium concentration following fertilization
triggers a number of events
• activation of cyclin-dependent kinases

Calcium waves are transient

• ions are rapidly pumped out of the cytosol and back into
the ER or extracellular space
 Ca ++
Binding Proteins
• Calcium affects a number of different types of cellular
effectors

Calcium ions can activate or inhibit enzymes and transport

system
• Calcium can also change the ionic permeability of
membranes

Calcium can also induce membrane fusion or alter

cytoskeletal structure and function
 Ca ++
Binding Proteins
• Calcium-Binding Proteins help calcium ions achieve its
various functions

Calmodulin
• Best studied example
• Universal

Same amino acid sequence from one end of the

eukaryotic spectrum to the other
• A molecule contains 4 binding sites for calcium

No sufficient affinity for calcium ions to bind in a

nonstimulated cell
 Calcium-Calmodulin Complex
• May bind to a protein kinase, a cyclic nucleotide
phosphodiesterase, ion channel, or the calcium-transport
system of the plasma membrane
• Also stimulate gene transcriptions through activation of
various protein kinases that phosphorylate transcription
factors
 Signal Transducers and
Activators of Transcription
(STATs)
• STAT proteins comprise a family of transcription
factors that become activated by tyrosine kinases
in the cytoplasm and then migrate to the nucleus
where they directly regulate gene expression.
 Structure-Function Relationships in STAT
Proteins

Fig 1. — Generic structure of a STAT protein illustrating common functional domain elements shared by
STAT family members. The sites of tyrosine (Y) and serine (S) phosphorylation are shown. SH2 = Src-
homology 2 domain, N = amino terminus, C = carboxyl terminus.

- a generalized diagram depicting the location of important

structural motifs common to most STAT family members.
- Each STAT molecule contains an Src-homology 2 (SH2) domain
Monomeric, inactive STAT proteins associate with each other to
form active dimers through a key phosphotyrosine (pY) residue,
which binds to the SH2 domain of another STAT monomer
Activating event in STAT signaling is tyrosine phosphorylation
N-terminal portion: DNA-binding domain
C-terminal portion: transactivation domain -serine residue
 Role of STATs in Normal Signal
Transduction

Fig 2. — Signal transduction pathways leading to STAT activation. Stimulation with growth factors or
cytokines at the cell surface results in receptor activation and subsequent tyrosine phosphorylation of
STATs. Phosphorylation of STATs induces dimerization and translocation to the nucleus, where STAT
dimers bind to specific STAT response elements and directly regulate gene expression. In contrast to
normal signaling, oncogenic PTKs constitutively activate STATs, leading to deregulated expression of
STAT-dependent genes. In some cases, but not all, JAK family tyrosine kinases are known to have a
role in STAT activation.