Beruflich Dokumente
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Biochemical Energetics
Go'
[C] [D]
+ RT ln
[A] [B]
For a reaction A + B C + D
[C] [D]
G = G' + RT ln
[A] [B]
At equilibrium
G = 0.
[C] [D]
G = G' + RT ln
[A] [B]
[C] [D]
K'eq, the ratio [C][D]/[A] = G' + RT ln [A] [B]
[C] [D]
G' = - RTln
[A] [B]
[ C] [ D]
defining K'eq =
[A] [B]
G' = - RT ln K'eq
Go' = RT ln K'eq
Variation of equilibrium constant with Go (25 oC)
K'eq
G '
kJ/mol
10
- 23
10
- 11
10 = 1
is at equilibrium
10
-2
+ 11
10
-4
+ 23
Energy coupling
A spontaneous reaction may drive a non-spontaneous
reaction.
Free energy changes of coupled reactions are additive.
A. Some enzyme-catalyzed reactions are interpretable as
two coupled half-reactions, one spontaneous and the
other non-spontaneous.
At the enzyme active site, the coupled reaction is
kinetically facilitated, while individual half-reactions
are prevented.
Free energy changes of half reactions may be summed,
to yield the free energy of the coupled reaction.
Go' = 31 kJ/mol
Coupled reaction:
ATP + glucose ADP + glucose-6-P Go' = 17 kJ/mol
The structure of the enzyme active site, from which H2O
is excluded, prevents the individual hydrolytic reactions,
while favoring the coupled reaction.
ADP + Pi
S2
S1
ATP
Side 1
Side 2
S1
S2
Side 1
Side 2
[S]2
+ Z F
G = R T ln
[S]1
R = gas constant, T = temperature, Z = charge on the ion,
F = Faraday constant, = voltage.
ADP + Pi
S2
S1
ATP
Side 1
Side 2
ADP + Pi
ADP + Pi
S2
S1
active
transport
ATP
H+2
H+1
ATP
synthesis
ATP
Two examples:
Active Transport: Spontaneous ATP hydrolysis
(negative G) is coupled to (drives) ion flux against a
gradient (positive G).
ATP synthesis: Spontaneous H+ flux (negative G) is
coupled to (drives) ATP synthesis (positive G).
ATP
adenosine triphosphate
O
-O
P
O-
O
O
P
O-
O
O
phosphoanhydride
bonds (~)
adenine
CH2
O-
OH
H
OH
ribose
ATP
adenosine triphosphate
O
-O
P
O-
O
O
P
O-
adenine
N
phosphoanhydride
bonds (~)
O
O
NH2
CH2
OH
OH
H
OH
ribose
AMP~P AMP + Pi
Alternatively:
AMP~P~P AMP + P~P
P~P 2 Pi
(PPi 2Pi)
NH2
Artificial ATP
analogs have
been designed
that are resistant
to cleavage of
the terminal
phosphate by
hydrolysis.
-O
O-
O-
O
O
CH2
OH
N
N
OH
H
OH
Example: AMPPNP.
Such analogs have been used to study the dependence of
coupled reactions on ATP hydrolysis.
In addition, they have made it possible to crystallize an
enzyme that catalyzes ATP hydrolysis with an ATP analog
at the active site.
Inorganic polyphosphate
Many organisms store energy as inorganic
polyphosphate, a chain of many phosphate residues
linked by phosphoanhydride bonds:
P~P~P~P~P...
Hydrolysis of Pi residues from polyphosphate may be
coupled to energy-dependent reactions.
Depending on the organism or cell type, inorganic
polyphosphate may have additional functions.
E.g., it may serve as a reservoir for Pi, a chelator of metal
ions, a buffer, or a regulator.
Phosphocreatine (creatine
phosphate), another
compound with a "high
energy" phosphate linkage,
is used in nerve & muscle
for storage of ~P bonds.
H
N
P
O
CH3
C
NH2
O
CH2
C
O
phosphocreatine
O
C
C
CH2
PEP
O
ADP ATP
OPO32
H+
C
C
OH
CH2
enolpyruvate
CH3
pyruvate
NH2
N
ester linkage
O
-O
P
O-
O
O
P
O-
O
O
CH2
O-
adenine
O
OH
H
OH
ribose
O
6 CH
OH
P
OH
O
H
OH
3
OH
H
1
H
2
OH
CH2
OH
glucose-6-phosphate
HO
OH
CH
CH2
O
O
glycerol-3-phosphate
Protein Kinase
OH + ATP
Protein
Protein
O + ADP
O
Pi
H2O
Protein Phosphatase
Go' of phosphate
hydrolysis, kJ/mol
Phosphoenolpyruvate (PEP)
Phosphocreatine
Pyrophosphate
Glucose-6-phosphate
Glycerol-3-phosphate
Some other
high energy
bonds:
O
Coenzyme A-SH + HO
O
Coenzyme A-S
+ H2O
O
C CH3
Coenzyme A-SH + HO
acetic acid
O
Coenzyme A-S C
acetyl-CoA
CH3 + H2O
SH
CH2
Coenzyme A includes
-mercaptoethylamine,
in amide linkage to the
carboxyl group of the B
vitamin pantothenate.
The hydroxyl of
pantothenate is in ester
linkage to a phosphate
of ADP-3'-phosphate.
The functional group is
the thiol (SH) of
-mercaptoethylamine.
-mercaptoethylamine
CH2
NH
C
CH2
pantothenate
CH2
NH
C
NH2
O
HO
H3C
CH3 O
H2C
P
O
O
O
CH2
ADP-3'-phosphate
Coenzyme A
H
OH
P
O
NH2
cAMP
N
H2
5' C 4'
O
O
O
H
H 3'
P
O
1'
2' H
OH
O-
Kinetics vs Thermodynamics:
A high activation energy barrier usually causes
hydrolysis of a high energy bond to be very slow in
the absence of an enzyme catalyst.
This kinetic stability is essential to the role of ATP and
other compounds with ~ bonds.
If ATP would rapidly hydrolyze in the absence of a
catalyst, it could not serve its important roles in energy
metabolism and phosphate transfer.
Phosphate is removed from ATP only when the reaction
is coupled via enzyme catalysis to some other reaction
useful to the cell, such as transport of an ion,
phosphorylation of glucose, or regulation of an enzyme
by phosphorylation of a serine residue.
C
H
H
H H
OH
H H
OH
NAD+, Nicotinamide
Adenine Dinucleotide,
is an electronacceptor
in catabolic pathways.
Nicotinamide
Adenine
Dinucleotide
+
N
CH2
nicotinamide
H
OH
OH
NH2
N
P
NH2
O
C
CH2
N
O
adenine
esterified to
Pi in NADP+
H
OH
OH
NAD+/NADH
H
O
C
+
N
NH2
2 e + H+
O
NH2
NAD+
NADH
dimethylisoalloxazine
H 3C
H 3C
H
C
C
H
FAD
C
C
C
C
O
C
2e +2H
NH
C
CH2
HC
OH
HC
OH
HC
OH O
H2C
P
O-
Adenine
O
O
P
O-
Ribose
H3C
H3C
H
C
C
H
C
C
FADH2
H
N
O
C
C
CH2
N
H
HC
OH
HC
OH
HC
OH O
H2C
P
O-
NH
C
Adenine
O
O
O-
Ribose