Biochemistry 2/e - Garrett & Grisham

Chapter 28
Metabolic Integration and
Unidirectionality of Pathways
to accompany
Biochemistry, 2/e
by
Reginald Garrett and Charles Grisham
All rights reserved. Requests for permission to make copies of any part of the work
should be mailed to: Permissions Department, Harcourt Brace & Company,
6277
Sea Harbor Drive, Orlando, Florida 32887-6777
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Outline

28.1 A Systems Analysis of Metabolism

28.2 Metabolic Stoichiometry
28.3 Unidirectionality
28.4 Metabolism in a Multicellular
Organism

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Systems Analysis of Metabolism

Catabolic and anabolic pathways, occurring

simultaneously, must act as a regulated, orderly,
responsive whole
See Figure 28.1 - catabolism, anabolism and
macromolecular synthesis
Just a few intermediates connect major systems sugar-Ps, alpha-keto acids, CoA derivs, and PEP
ATP & NADPH couple catabolism & anabolism
Phototrophs also have photosynthesis and CO2
fixation systems
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

28.2 Metabolic Stoichiometry

Three types of stoichiometry in biological
systems
Reaction stoichiometry - the number of each
kind of atom in a reaction
Obligate coupling stoichiometry - the
required coupling of electron carriers
Evolved coupling stoichiometry - the number
of ATP molecules that pathways have
evolved to consume or produce - a number
that is a compromise, as we shall see
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

The Significance of 38 ATPs

The "ATP stoichiometry" has a large effect on the
Keq of a reaction
Consider the Keq for glucose oxidation (page 932)
If 38 ATP are produced, cellular G is -967 kJ/mol
and Keq = 10170, a very large number!
If G = 0, 58 ATP could be made, but the reaction
would come to equilibrium with only half as much
glucose oxidized as we could have had
So the number of 38 is a compromise!

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Significance of large Keq

The more ATP obtained, the lower the
equilibrium constant, and the higher the level
of glucose required
If [glucose] is below this value, it won't be
effectively utilized
Large Keq means that this threshold level of
glucose will be be very low
Large Keq also means that the reaction will be
far from equilibrium and can thus be regulated
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

The ATP Equivalent

What is the "coupling coefficient" for ATP produced or
consumed?
Coupling coefficient is the moles of ATP produced
or consumed per mole of substrate converted (or
product formed)
Cellular oxidation of glucose has a coupling
coefficient of 30-38 (depending on cell type)
Hexokinase has a coupling coefficient of -1
Pyruvate kinase (in glycolysis) has a coupling
coefficient of +1
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

The ATP Value of NADH vs

NADPH
The ATP value of NADH is 2.5-3
The ATP value of NADPH is higher
NADPH carries electrons from catabolic
pathways to biosynthetic processes
[NADPH]>[NADP+] so NADPH/NADP+ is a
better e- donating system than NADH/NAD
So NADPH is worth 3.5-4 ATP!

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Nature of the ATP Equivalent

A different perspective
G for ATP hydrolysis says that at equilibrium
the concentrations of ADP and Pi should be
vastly greater than that of ATP
However, a cell where this is true is dead
Kinetic controls over catabolic pathways ensure
that the [ATP]/[ADP][Pi] ratio stays very high
This allows ATP hydrolysis to serve as the
driving force for nearly all biochemical
processes
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Solvent Capacity of the Cell

The capacity to keep all metabolites solvated

What is the role of ATP in solvent capacity?
Consider phosphorylation of glucose
If done by Pi, the concentration of Pi would have
to be 2700 M
However, using ATP, and if [ATP] and [ADP] are
equal, [G-6-P]/[G] is maintained at 850
ATP, an activated form of phosphate, makes it
possible for cell to carry out reactions while
keeping concentrations of metabolites low
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Substrate Cycles
If ATP c.c. for a reaction in one direction differs from
c.c. in the other, the reactions can form a
substrate cycle
See Figure 28.2
The point is not that ATP can be consumed by
cycling
But rather that the difference in c.c. permits both
reactions (pathways) to be thermodynamically
favorable at all times
Allosteric effectors can thus choose the direction!
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Unidirectionality of Pathways
A "secret" role of ATP in metabolism
Both directions of any pair of opposing
pathways must be favorable, so that
allosteric effectors can control the direction
effectively
The ATP coupling coefficient for any such
sequence has evolved so that the overall
equilibrium for the conversion is highly
favorable
See Figure 28.4 for an illustration!
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Energy Charge
Adenylates provide phosphoryl groups to
drive thermodynamically unfavorable
reactions
Energy charge is an index of how fully
charged adenylates are with phosphoric
anhydrides
If [ATP] is high, E.C.1.0
If [ATP] is low, E.C. 0
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Fueling the Brain

Brain has very high metabolism but has
no fuel reserves
This means brain needs a constant
supply of glucose
In fasting conditions, brain can use hydroxybutyrate (from fatty acids),
converting it to acetyl-CoA in TCA
This allows brain to use fat as fuel!
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Creatine Kinase in Muscle

Muscles must be prepared for rapid provision
of energy
Creatine kinase and phosphocreatine act as a
buffer system, providing additional ATP for
contraction
Glycogen provides additional energy, releasing
glucose for glycolysis
Glycolysis rapidly lowers pH, causing muscle
fatigue
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Muscle Protein Degradation

During fasting or high activity, amino
acids degrade to pyruvate, which can be
transaminated to alanine
Alanine circulates to liver, where it is
converted back to pyruvate - food for
gluconeogenesis
This is a fuel of last resort for the fasting
or exhausted organism
Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company

Biochemistry 2/e - Garrett & Grisham

Copyright 1999 by Harcourt Brace & Company