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Chapter 15:Metabolism:
Basic Concepts and Design
Copyright © 2007 by W. H. Freeman and Company
Berg • Tymoczko • Stryer
BiochemistrySixth Edition
Roadmapof
Metabolic Pathways
Metabolism
Metabolism – reactions occurring in a living system that produce and consume the energy needed for the organism to exist.
•Metabolic pathways.
•Metabolic reactions.
•High Energy bonds in compounds.
•Thermodynamics of reactions.
Metabolism
• Metabolism - the entire network of chemical reactions carried out by living cells
• Metabolites - small molecule intermediates in the degradation and synthesis of biopolymers
• Catabolic reactions - degrade biomolecules to create smaller molecules and energy
• Anabolic reactions - synthesize biomolecules for cell maintenance, growth and reproduction
Catabolism and Anabolism
Catabolism Anabolism
degradative synthetic
oxidative reductive
energy producing energy requiring (exergonic) (endergonic)
makes pool molecules uses pool molecules
produces NADH & uses NADPH almost NADPH exclusively
Energy Overview
Energy distribution
1/3 2/3 nutrients ----> pool molecules ----> CO2, H2O,
NH3
biomolecules
Pathways
• Metabolism includes all enzyme catalyzed reactions
• Metabolism can be subdivided into various areas: hexose shunt, electron transport, etc.
• The metabolism of the four major groups of biomolecules will be considered:
CarbohydratesLipidsAmino AcidsNucleotides
Pathways
• Multiple-step pathways permit control of energy input and output
• Catabolic multi-step pathways provide energy in smaller stepwise amounts)
• Each enzyme in a multi-step pathway usually catalyzes only one single step in the pathway
• Control points occur in multistep pathways
Regulation
• Metabolism is highly regulated to permit organisms to respond to changing conditions
• Most pathways are irreversible
• Flux - flow of material through a metabolic pathway which depends upon:
(1) Supply of substrates(2) Removal of products(3) Pathway enzyme activities
Levels of Regulation
1. Direct regulation at the enzyme level (covalent or non-covalent).
2. Regulation via external communication (hormonal).
3. Regulation at the gene level (induction/repression).
Direct Regulation
• Feedback Inhibition: The product of a pathway controls its own synthesis by inhibiting an earlier step (the first step or the “committed” step in the pathway) .
• Feed-forward Activation: A metabolite early in the pathway activates an enzyme that appears later.
• Interconvertible enzyme activity can be rapidly and reversibly altered by covalent modification. E.g. protein kinases and protein phosphatases.
GlucoseMetabolism
Breakdown to small molecules and energy.
Metabolite
Needed for formation of glycerol basedphospholipids and to run theglycerol-Pshuttle.
Adenosine Nucleotides
Components of an energy system.
ATPAn energy carrier considered to becommon energy currency in a cell
Driving Forces behind the Energy of ATP Hydrolysis
1. Resonance energy of reactants vs products.
2. Charge repulsion of oxygens.
3. Number of charges on oxygens.
4. Solvation of reactants vs products.
5. Entropy – number of reactant vs product molecules.
Phosphate Resonance
pKas of phosphoric acid:2.1, 6.9 and 12.3
Other HighEnergy
Molecules
Go'of Hydrolysis
ATP
Use
Synthesis
Oxidation States
Oxidation of triacylglycerols affords more energy than do carbohydrates.
Sources of Energy
Biological Redox Energy
• Electron Transport System (ETS) moves electrons from reduced coenzymes toward O2
• This produces a proton gradient and a transmembrane potential
• Oxidative Phosphorylation is the process by which the potential is coupled to the reaction:
ADP + Pi ATP
NAD+ Oxidizes GAP
NADH carries electrons to the ETS.
Substrate Level Phosphorylation
Substrate Level Phophoryation occurs When ATP is formed in a metabolic reaction.
Free Energy of Coupled Reactions
ADP + Pi --- >ATP
1,3-bisphosphoglycerate --- > 3-phosphoglycerate + Pi
1,3-bisphosphoglycerate + ADP ---- >3-phosphoglycerate + ATP
Go' = -49.4 kJ/mol
Go' = +30.5 kJ/mol
Go' = -18.9 kJ/mol
AerobicOxidation
Oxidative phosphorylation does not occur without electron transport.
MitochondriaOxidation and electron transport
Oxidative phosphorylation
NAD+
NicotinamideNucleotide
AMPR = -PO3= for NADP+
AMP = AdenineNucleotide
A two electron transfer agent
Oxidation by NAD+
This side is the “A” face of the nicotinamide ring, the back side is the “B” face.
Oxidation by NAD+
A typically NAD+ oxidation is -OH to C=O
FAD
FMN = Flavin Mononucleotidein blue
AMP in black
A one electron transfer agent
Note that this is ribitol.
Oxidation by FAD
FAD and FMN also accept two electrons but these enter the isoalloxazine ring one at a time.
Oxidation by FAD
A typically FAD oxidation is -CH2-CH2- to -CH=CH-
Oxidized and Reduced Forms
This is an isoalloxazine ring system
Coenzyme A
An acyl transfer agent (forms a thioester)
Note -PO3= on 3' of ribose
Thioesters
Carriers and Coenzymes
Review of G Equations
• For the reaction: A + B C + D
• At standard state: All conc. are 1 M or 1 atm except [H+] and under these conditions:
G = Go'
G = Go' + RT ln([C][D]/[A][B])
Review of G Equations
• For the reaction: A + B C + D
• At equilibrium: Keq = [C]eq[D]eq/[A]eq[B]eq and G = 0, therefore:
Go' = -RT ln Keq
Go' = -nEo'F
• For an oxidation-reduction reaction:
(#e transferred)(cell potential)(Faraday’s const.)
Krebs Cycle Oxidations
Also, there are two oxidative decarboxylations in the Kreb’s Cycle (citric acid cycle).
Free Energy of a Redox Reaction
Oxidation Half-reaction: Half-Cell PotentialMalate ---- > Oxaloacetate + 2 e + 2 H+ Eo' = +0.166 v
Reduction Half-reaction:NAD+ + 2 e + 2 H+ ---- > NADH + H+ Eo' = -0.32 v
Cell Reaction :Malate + NAD+ ---- > Oxaloacetate + NADH + H+
Cell Potential: Eo' = -0.154 v
A cell reaction must contain an oxidation half-reaction and a reduction half-reaction to equate electron flow.
Free Energy of a Redox Reaction
Go' = -nEo'F
= -(2)(-0.154)(96480)= +29700 J/mol= +29.7 kJ/mol
The equilibrium of this redox reaction lies far to the left. Cellular concentrations of the metabolites must be such that the overall G is negative in order for the reaction to proceed as written on the previous slide. For a redox reaction to proceed spontaneously, the cell potential must be positive.
Free Energy of a Redox Reaction
Which reactant is oxidized ?
Which reactant is reduced ?
Which reactant is the oxidizing agent ?
Which reactant is the reducing agent ?
Malate + NAD+ ---- > Oxaloacetate + NADH + H+
Malate
NAD+
NAD+
Malate
Reaction Types in Metabolism
Ligation with ATP
Isomerization
Group Transfer
Hydrolysis
Cleavage to form a Double Bond
Cleavage to form a Double Bond
Energy Charge of a Cell
ATP + ½ ADP Energy Charge = ------------------------- ATP + ADP + AMP
Limits are 0 and 1.0
If all is ATP, the energy charge = 1 If all is AMP, the energy charge = 0
ATP can be regenerated using adenylate kinase (this is a nucleoside monophosphate kinase):
2 ADP <===> ATP + AMP
Rate vs Energy Charge
Other ATP uses
ATP can also be used to make other NTPs with nominal energy exchange using a nucleoside diphosphate kinase.
ATP + NDP <===> ADP + NTP
Other involvement of ATP:
1. Phosphate transfer to make high energy bond:Glutamine synthesis uses P from ATP
Glu + ATP —> γ-PGlu + ADP, then NH3 displaces P to give Gln
Other ATP uses
2. PEP transfers P to make ATP:
Enol-P (PEP) + ADP —> Pyr + ATP
3. Nucleotide transfer to make high energy bond:AMP from ATP combines with a fatty acid in making AcylSCoA catalyzed by acylSCoA synthetase (acyl thiokinase) during fatty acid activation.
FA + ATP —> acyl-AMP + PPi, then CoASH displaces AMP to give acyl-SCoA
Effect of H+ on Keq
pyruvate + NADH + H+ ----> lactate + NAD+
[lactate][NAD+]Keq = -------------------------------
[pyruvate][NADH][H+]
[lactate][NAD+]Keq' = Kapp = -------------------------
[pyruvate][NADH]
so, Keq' = Keq (H+), where H+ is a reactant.similarly, Keq' = Keq /(H+), where H+ is a product.
FAD vs FAD-flavoprotein
Electrons from succinate:FADH2 + CoQ < === > FAD + CoQH2
Go' for free FAD in solution:
FAD + 2 H+ + 2 e- <===> FADH2 Eo' = -0.22vCoQ + 2 H+ + 2 e- <===> CoQH2 Eo' = +0.10v
net FADH2 + CoQ <===> FAD + CoQH2 Eo' = +0.32v
Go' = -nEo'F = -61.7 kJ/mol
FAD vs FAD-flavoprotein
CoQ + FADH2 < === > CoQH2 + FAD
Go' for FAD in a flavoprotein:
FAD + 2 H+ + 2 e- <===> FADH2 Eo' = 0.00vCoQ + 2 H+ + 2 e- <===> CoQH2 Eo' = +0.10v
net FADH2 + CoQ <===> FAD + CoQH2 Eo' = +0.10v
Go' = -nEo'F = -19.3 kJ/mol
This represents a difference in Go' of about 42 kJ/mol.
Table of Reduction Potentials
End of Chapter 15
Copyright © 2007 by W. H. Freeman and Company
Berg • Tymoczko • Stryer
BiochemistrySixth Edition