Energy is not Cycled

CO
2
Glucose
CO
2

Fat, muscle, etc.
Anabolism
Energy (light) Energy (heat)
Energy (heat)
Catabolism
Catabolism Drives Analobolism
Catabolism
Breakdown of nutrients (e.g.
glucose)
Releases energy for anabolic
reactions
Releases heat

Anabolism (Biosynthesis)
Synthesis of macromolecules
Requires energy
Releases heat

Intermediary Metabolism
Metabolic pathways involving low
molecular weight (<1000)
compounds


Catabolism Driving Anabolism
Separation of Anabolic and Metabolic
Pathways
Unique Enzymes

Glucose to G-6-P via glucokinase (catabolic) (uses ATP)
G-6-P to glucose by G-6-P phosphatase (uses Pi)
Differential regulation (usually one off, the other on)
Glucose + ATP
Glucose-6-Phosphate
Glucokinase
Phosphate + Glucose
G-6-P
Phosphatase
Separation of Anabolic and Meabolic
Pathways

Cellular Compartmentalization
Fatty acid catabolism in mitochondria
Fatty acid synthesis in cytoplasm
Different concentrations of products, reactants and regulators
Unique Cofactors
NAD/NADH for catabolism
NADP/NADPH for anabolism

When it goes wrong .
Classes of Biochemical Reaction
Oxidation-reduction
Lactate dehydrogenase
Cleavage of carbon bonds
Adol condensations (aldose)
Claison condensations (citrate synthase)
Decarboxylations (acetoacetate decarboxylase)
Internal rearrangements, isomerizations and eliminations
Phosphohexose Isomerase
Group transfers (eg acyl,glucosyl, phosphoryl)
Hexokinase
Free radical reactions
Ribonucleotide reductase
Oxidation-Reduction
Oxidation-Reduction
Dehydrogenases
Dehydrogenations - loss of 2 electrons & 2 hydride ions
Oxidases
Oxygen becomes bonded to carbon
Oxygenases
Oxidases that use molecular oxygen
Oxidation
Reduction
Phosphate and PhosphateTransfer
Transient Intermediate
Nucleophilic Attack (Glucokinase)
Consider a Typical Enzymatic Reaction
For the reaction:
At equilibrium:
Where K
eq
= the equilibrium constant
[A] = the molar concentration of A etc.
----------------------------------------------------------------------
K'
eq
pH 7 (10
-7
M H
+
) 55.5 M H
2
O
A BC D
K
eq

[C][D]
[A][B]
The reaction is reversible
Forward and backward reactions occur at the same time
The rate of each reaction is dependent on the concentration
of reactants
As A and B are used up, the forward rate decreases, C and D
increase and the rate of the reverse reaction increases
At the steady state, the forward and backward reaction rates
are the same
For the forward reaction;
A and B are substrates
C and D are products
Consider a Typical Enzymatic Reaction
For the reaction:
At equilibrium:
Where K
eq
= the equilibrium constant
[A] = the molar concentration of A etc.
----------------------------------------------------------------------
K'
eq
pH 7 (10
-7
M H
+
) 55.5 M H
2
O
A BC D
K
eq

[C][D]
[A][B]
Forward Rate
Proportional to [A][B] = k
1
[A][B]

Reverse Rate
Proportional to [C][D] = k
2
[C][D]

At Equilibrium: forward rate = reverse rate




k
1
and k
2

are
constants
k
1
[C][D]
k
2
[A][B]

=
k1[A][B] = k2[C][D]
Kd [C][D]
[A][B]

=
Equilibrium Constant
For the reaction:
At equilibrium:
Where K
eq
= the equilibrium constant
[A] = the molar concentration of A etc.
----------------------------------------------------------------------
K'
eq
pH 7 (10
-7
M H
+
) 55.5 M H
2
O
A BC D
K
eq

[C][D]
[A][B]
Equilibrium Constant
All enzymatic
reactions are
reversible
ABC D
K
eq

[C][D]
[A][B]
Enzymatic Reactions are reversible
A or B
Forward
C or D
A or B
C or D
Backwards
}
}
At dynamic
equilibrium
Change in Gibbs Free Energy G
The free energy change drives a reaction
A negative G drives the reaction forward (as written)
A positive G drives the reaction backwards (as written)
The G depends on the concentrations of reactants and
products, temperature and pressure
Standard G (G)
One molar reactants and products, 298 K (25 C), 1 atm
Standard Biological G (G)
As G + pH 7 (10
-7
M H
+
), 55.5 M H
2
O, 1 mM Mg
2+

The standard free energy change of a reaction cannot
used to reliably predict the net direction of a reaction in
vivo since the reactants and products are not at 1 M
Enzymes Lower the Activation Energy
Enzymes work by lowering the activation energy

Therefore:

Enzymes increase the dynamics of a reaction
Enzymes may increase the net rate of product formation
Enzymes do not change the K
eq

Free Energy Change
G = free energy change
G = standard G
G' = biological standard G
1 M, 298 K, 1 atm
10
-7
M H
+
, 55.5 M H
2
O
ABC D
1 mM Mg
2+

Relationship between G and K
eq

For the reaction
From thermodynamic principles:
At equilibrium
Therefore:
R= the gas const ant
T= the t emperature in Kelvin
G G
0'
RTln
[C][D]
[A][B]
A BC D
G
o'
RTlnK
eq
'
G 0
Law of Mass Action
G G
0'
RTln
[C][D]
[A][B]
G0 G
0'
RTln
[C][D]
[A][B]
At equilibrium
ABC D
Remove some of the products C and D
G is zero:
G becomes negative
G0 G
0'
RTln
[C][D]
[A][B]
Remove some of the substrates A and B
G becomes positive
G0 G
0'
RTln
[C][D]
[A][B]
A + B C + D
A + B C + D
Forwards
Backwards
K
eq

[C][D]
[A][B]
Gs are Additive
Glucose + P
i
G-6-P + H
2
0
ATP + H
2
0 ADP + P
i

Since only starting and final states are considered:
Glucose + ATP G-6-P + ADP
+13.8 kJ/mol
G'
-30.5 kJ/mol
-16.7 kJ/mol
The hydrolysis of ATP is used to drive the reaction forward
Consider the reaction:
Energetically unfavorable reactions are coupled to favorable ones
Keqs are Multiplicative
Glucose + P
i
G-6-P + H
2
0
ATP + H
2
0 ADP + P
i

Since only starting and final states are considered:
Glucose + ATP G-6-P + ADP
3.9 x10
-3
M
Keq
2.0 x 10
5
M
7.8 x 10
2
M
The hydrolysis of ATP is used to drive the reaction forward
Consider the reaction:
Energetically unfavorable reactions are coupled to favorable ones
Overall G = G
1
+ G
2
where G
1
= RTln Keq
1

Overall G = RTln Keq1 + RTln Keq2 (adding logs = multiplication)
Operational Gs
ATP + H20 ADP + Pi
G'
-30.5 kJ/mol
Consider the reaction:
Observed versus biological standard Gs
Conditions vary, but in general the true G of ATP
hydrolysis in vivo is more negative than standard G
In human erythrocytes at 25 C
[ADP] = 0.25 x 10
-3
[Pi] = 1.65 x 10
-3

[ATP] = 2.25 x 10
-3

Gp = G + RT ln [ADP][Pi]
[ATP]
Gp = -30.5 + 21 kJ/mol
G of ATP hydrolysis = 51.5 kJ/mol
Metabolic Pathways & Law of Mass Action
Products are moved through pathways by:
Increasing the level of substrate
Decreasing the level of product
Especially useful for energetically unfavorable
reactions

ABC D
K
eq

[C][D]
[A][B]
G G
0'
RTln
[C][D]
[A][B]
Relationship Between K
eq
and G
Keq G (kJ/mol)
0.001 17.1
0.01 11.4
0.1 5.7
1 0.0
10 -5.7
100 -11.4
1,000 -17.1
Why is G 0 when K
eq
is 1?
ATP
Adenosine
ATP
AMP
ADP
Phosphoester
bonds
Phosphoanhydride
bonds
a b g
ATP Hydrolysis
Hydrolysis with relief of charge repulsion
Resonance stabilization Ionization
ATP Forms a Complex With Mg
2+

ATP is a High-Energy Compound
Hydrolysis of Phosphoenolpyruvate
Tautomerization
G = -16 kJ/mol G = -46 kJ/mol
PEP
3-
+ H
2
O Pyruvate + P
i
2-

G = -16 kJ/mol
1,3-BPG Hydrolysis
H
+

1,3-Bisphosphoglycerate
4-
+ H
2
O



3-phosphoglycerate
3-
P
i
2-
+ H
+

G = -49.3 kJ/mol
ionization
Resonance Stabilization and Ionization
Phosphocreatine Hydrolysis
Resonance Stabilization
Transphosphorylations
Phosphocreatine is an Energy Reservoir
ADP + phosphocreatine ATP + creatine G = -12.5 kJ/mol
2ADP ATP + AMP G = ~ 0
ATP + NDP ADP + NTP G = ~ 0
Creatine Kinase
Other Transphosphorylations
Adenylate Kinase
Nucleoside diphosphate Kinase
Acetyl-CoA Hydrolysis
Resonance Stabilization and Ionization
Thioesters Versus Oxygen Esters
No Resonance Stabilization in Thioesters
Comparison of Gs of Hydrolysis
Standard Free Energies of Hydrolysis
G kJ/mol
(approx)
Phospoenolpyruvate - 62
1,3-bisphoshoglycerate to 3-BPG - 49
Phosphocreatine - 43
ATP to ADP
ATP to AMP + PP
i

ADP to AMP


PP
i

AcetylCoA - 31
Hexose phosphates - 13 to - 21
AMP - 14
Gycerol-1-phosphate - 9.2
{
~ - 31
ATP Coupling is Multi-Step
Glutamine Synthetase
Na+K+ ATPase
ATP Hydrolysis Powers the Pump
ATPase Pumping Mechanism
Phosphorylation changes the conformation of the pump
Palmitoyl-CoA Synthesis
AMP is conjugated to palmitate
RNA Elongation
Two High-Energy Bonds are Used
Oxidation
Food for Work
Comparison to an electrical motor
Battery
Electric
Motor
WORK
e.g. movement
Food Mitochondria WORK
e.g. muscle use
Electrons
Physical
Coupling
Electrons ATP
Electromotif Force (emf)
Proton Motive Force
Redox Pair
Fe
2+
Cu
2+
Fe
3+
Cu
+
+ +
Fe
2+
Cu
2+
Fe
3+
Cu
+
+
+
e
-
e
-
Reducing agent
(reductant,
Oxidizing agent
(oxidant, electron
acceptor)
electron donor)
Oxidation -
Reduction
Ferrous Ferric Cupric Cuprous
Oxidation
loss of electrons
Reduction
gain of electrons
Transfer of Electrons in Organic Compounds
Oxidation of a reducing sugar by cupric ions
C R
O
H
+
4OH
-
+
2Cu
2+
Cu
2
O + 2H
2
O
C R
O
OH
+
C R
O
H
+ 2OH
-
+
H
2
O
C R
O
OH
+
2OH
-
+ 2Cu
2+
Cu
2
O H
2
O
+ + 2e
-
2e
-
Oxidation-Reduction
Oxidation - loss of electrons
Reduction - gain of electrons
cupric ion
Transfer of Electrons in Organic Compounds
Oxidation of a reducing sugar by cupric ions

C R
O
H
+
4OH
-
+
2Cu
2+
Cu
2
O + 2H
2
O
C R
O
OH
+
C R
O
H
+ 2OH
-
+
H
2
O
C R
O
OH
+
2OH
-
+ 2Cu
2+
Cu
2
O H
2
O
+ + 2e
-
2e
-
Oxidation-Reduction
2OH
-
give H
2
O plus O
2-

2Cu
2+
+ 2e
-
give 2Cu
+

2Cu
+
+ O
2-
give Cu
2
O
OH replaces H yields 1e
-

H combines with OH
-
to give H
2
O plus 1e
-
Oxidation States of Carbon
Ownership of Electrons H< C < S < N < O
E
l
e
c
t
r
o
n
s

b
e
l
o
n
g
i
n
g

t
o

c
a
r
b
o
n

Methods of Electron Flow
Fe
2+
+ Cu
2
+ Fe
3+
+ Cu+
Direct Electron Transfer
Use of Hydride Ions
Use of Hydrogen Atoms
Direct Oxidation by Oxygen to Give
4 Methods of Electron Flow
a Covalently Bound Oxygen
AH
2
+ B A + BH
2
AH
2 A + 2H
+
2e
-
NAD
+
+ H
-
+ H
+
e
-
NADH + H+
NAD
+
+ AH
2
NADH + H
+
R-CH
3
+
1
/
2
O
2 R-CH
2
-OH
Electron acceptor
Electron acceptor
Electron acceptor
Nernst Equation
E = E
0
+
RT
nF
ln
[electron acceptor]
[electron donor]
__ _______________
E = E
0
+
n
ln
[electron acceptor]
[electron donor]
_______________
0.026 V
NERNST EQUATION
n =no. of electrons/molecule
F = Faraday Const.
T= temp (K)
R = gas constant
Relates Standard Reduction Potential to True Reduction Potential
E is the biological reduction
potential (pH 7)
Relationship of G to E
G = -n F E
or
G
0
' = -n F E
0
'
F = Faraday Const.
The Hardest Question on the Exam
G = -n F E
or
G
0
' = -n F E
0
'
F = Faraday Const.
Given E for two reactions, predict the
favorable direction the reaction; what
becomes oxidized and what reduced
Preparation
Work through the example in Lehninger pages 510/1
NAD+ Accepts 2 Electrons
Transferred as a Hydride Ion (H-)
Examples of NAD specificity
NADH Absorbs at 340 nm
Can be used to measure concentration
Pellagra
Dermatitis, diarrhea, dementia, death
Maize is deficient in tryptophan and niacin
Alcoholics have reduced absorption of niacin
QuickTime and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
Pellagra, a history
Once common in the Southern
US, pellagra was thought to be
contagious
1914 - Dr Joseph Goldberg went
to the S. to find a cure
Came to believe it was related to
nutrition
Controlled trials in orphanages
and a convict camp (induced
pellagra using a restricted diet)
In 1937 vitamin B3 was identified
Native Americans did not suffer
from pellagra because they treated
corn with lime which made niacin
available
QuickTime and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
Flavin adenine dinucleotide (FAD)
Accepts One or Two Electrons
Flavoproteins