Biological oxidation (part - III) Oxidative PhosphorylationAshok Katta
Biological oxidation (part - III) Oxidative Phosphorylation
- Mechanism of Oxidative Phosphorylation
-- Chemiosmotic theory
-P:O Ratio
Substrate Level Phosphorylation
Shuttle Systems for Oxidation of Extramitochondrial NADH
Substrate level phosphorylation and it's mechanism || Biochemistry || B Pharmacy || Project || slideshare || biology || chemistry
*images use in this ppt is only for educational purpose
In this presentation, i tell about substrate level phosphorylation
Phosphorylation involves the transfer of phosphate
group from one compound to other.
➢ Substrate level phosphorylation is a direct
phosphorylation of ADP with a phosphatase group by
using the energy obtain from a coupled reaction.
➢ Occurs in cytoplasm ( glycolysis – due to aerobic and
anaerobic condition) and in mitochondrial matrix ( krebs
cycle – anaerobic condition)
content:-
1. Introduction
2. Fermentation pathway
3. Production of some other foods & industrial chemical by use of fermentation
4. Energetics of fermentation
5. Summary
Biological oxidation (part - III) Oxidative PhosphorylationAshok Katta
Biological oxidation (part - III) Oxidative Phosphorylation
- Mechanism of Oxidative Phosphorylation
-- Chemiosmotic theory
-P:O Ratio
Substrate Level Phosphorylation
Shuttle Systems for Oxidation of Extramitochondrial NADH
Substrate level phosphorylation and it's mechanism || Biochemistry || B Pharmacy || Project || slideshare || biology || chemistry
*images use in this ppt is only for educational purpose
In this presentation, i tell about substrate level phosphorylation
Phosphorylation involves the transfer of phosphate
group from one compound to other.
➢ Substrate level phosphorylation is a direct
phosphorylation of ADP with a phosphatase group by
using the energy obtain from a coupled reaction.
➢ Occurs in cytoplasm ( glycolysis – due to aerobic and
anaerobic condition) and in mitochondrial matrix ( krebs
cycle – anaerobic condition)
content:-
1. Introduction
2. Fermentation pathway
3. Production of some other foods & industrial chemical by use of fermentation
4. Energetics of fermentation
5. Summary
Pentose phosphate pathway is also called Hexose monophosphate pathway/ HMP shunt/ Phosphogluconate pathway.
It is an alternative route for the metabolism of glucose.
It is more complex pathway than glycolysis.
It is more anabolic in nature.
It takesplace in cytosol.
The tissues such as liver, adipose tissue, adrenal gland, erythrocytes,testes and lactating mammary gland are highly active in HMP shunt.
It concern with the biosynthesis of NADPH and pentoses.
Complete Set of Metabolism of Carbohydrate in that second chapter, glycolysis.
This presentation covers complete glycolysis pathway with step wise animated reactions and it includes clinical aspects also. This presentation is good for MBBS students.
ATP synthase—also called FoF1 ATPase is the universal protein that terminates oxidative phosphorylation by synthesizing ATP from ADP and phosphate.
ATP Synthase is one of the most important enzymes found in the mitochondria of cells
Pentose phosphate pathway is also called Hexose monophosphate pathway/ HMP shunt/ Phosphogluconate pathway.
It is an alternative route for the metabolism of glucose.
It is more complex pathway than glycolysis.
It is more anabolic in nature.
It takesplace in cytosol.
The tissues such as liver, adipose tissue, adrenal gland, erythrocytes,testes and lactating mammary gland are highly active in HMP shunt.
It concern with the biosynthesis of NADPH and pentoses.
Complete Set of Metabolism of Carbohydrate in that second chapter, glycolysis.
This presentation covers complete glycolysis pathway with step wise animated reactions and it includes clinical aspects also. This presentation is good for MBBS students.
ATP synthase—also called FoF1 ATPase is the universal protein that terminates oxidative phosphorylation by synthesizing ATP from ADP and phosphate.
ATP Synthase is one of the most important enzymes found in the mitochondria of cells
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3. Oxidative PhosphorylationOxidative Phosphorylation
Electron transfer through the respiratory chain
releases about 200 kJ per “mole” of electron pairs.
This energy is conserved as a proton-motive force.
The formation of a mole of ATP from ADP and Pi
requires about 30 kJ.
How is a concentration gradient of protons
transformed to generate ATP?
4. Chemiosmotic TheoryChemiosmotic Theory
Proposed by Peter
Mitchell
The proton-motive
force, inherent in the
proton gradient,
drives the synthesis
of ATP as protons
flow passively back
into the matrix
through a protein
pore associated with
ATP synthase
5. Electron transport and ATP synthesis areElectron transport and ATP synthesis are
coupledcoupled
This can be demonstrated
when isolated mitochondria
are incubated and O2
consumption and ATP
synthesis measured.
Inhibitors of the passage of
electrons to O2 (e.g. cyanide,
carbon monoxide, and
antimycin A) block ATP
synthesis.
6. Conversely, inhibition of ATP synthesisConversely, inhibition of ATP synthesis
blocks electron transportblocks electron transport
without ADP, ATP is not made, but also, oxygen is not consumed
the toxic antibiotics oligomycin or venturicidin bind to the ATP
synthase and inhibit ATP synthesis and also O2 consumption
these toxins do not interact with electron carriers
therefore, inhibition of the ATPase blocks electron transport
electron transport and ATP synthesis are obligately coupled: neither
reaction occurs without the other
some compounds “uncouple” oxidation from phosphorylation, e.g.
dinitrophenol- dissipates the proton gradient
7. Why does electron transport depend onWhy does electron transport depend on
ATP synthesis?ATP synthesis?
When ATP synthase is inhibited, no path exists for the
return of protons to the matrix.
The continued extrusion of protons by the respiratory
chain generates a large proton gradient - the energy
required to pump protons against this gradient equals
or exceeds the energy provided by electron transfer.
At this point, electron flow stops.
8. ATP Synthase has two functional domainsATP Synthase has two functional domains
ATP synthase is a F-
type ATPase
Two distinct
components:
- F1 is a peripheral
membrane protein
that catalyzes the rxn
ADP + Pi ATP
- F0 is integral to the
membrane and
contains a proton
pore
9. ATP is stabilized relative to ADP on theATP is stabilized relative to ADP on the
surface of Fsurface of F11
On the enzyme’s surface, the reaction
ADP + Pi ATP + H2O
is readily reversible - the free energy change for ATP synthesis
is close to zero.
labeling experiments have shown that the terminal
pyrophosphate bond of ATP is cleaved and re-formed
repeatedly before Pi leaves the enzyme surface.
ATP synthase binds ATP tightly, and the free energy of
enzyme-bound ATP is close to that of ADP + Pi - on the
enzyme surface, the reaction is reversible and the equilibrium
constant close to 1.
The energy consuming step is release of the bound ATP, and
this is provided by the proton-motive force.
10. In a typical enzyme-catalyzed reaction, reaching the transition
state (F) between substrate and product is the major energy
barrier. For ATP synthase, release, not formation, of ATP is the
major energy barrier
11. For the continued synthesis
of ATP in this way, ATP
synthase must cycle
between a form that binds
ATP very tightly and a form
that releases ATP.
As protons flow, the cylinder
(c12 subunits) and shaft (γ
subunit) rotate, and the β
subunits of F1, which are
fixed in place relative to the
membrane, change
conformation as the γ
subunit associates with each
in turn
12. Binding-change model for ATP synthaseBinding-change model for ATP synthase
The F1 complex has three non-
equivalent adenine nucleotide
binding sites, one for each pair of
α and β subunits. Rotation of the
central shaft converts the sites as
follows:
β-ATP→ β-empty, ATP dissociates
β-ADP→ β-ATP, promotes ATP formation
β-empty→ β-ADP, loosely binds ADP + Pi
13. Electrons, protons, and ATP- what’s theElectrons, protons, and ATP- what’s the
stoichiometry?stoichiometry?
How many protons are pumped outward by electron transfer
from one NADH to O2?
Consensus values for protons pumped:
10 for NADH
6 for succinate
How many protons must flow inward through the F0 F1
complex to drive the synthesis of one ATP?
Consensus value for number of protons for one ATP = 4
P/O values (# NADH’s or succinate/ATP)
10/4 = 2.5 for NADH
6/4 = 1.5 for succinate
14. Complete oxidation of a molecule of glucose to CO2 yields 30 to
32 ATP molecules.
Overall efficiency = 68%
15. Cyanide PoisoningCyanide Poisoning
A. 22 year old comatose man had odor of almonds and
severe metabolic acidosis.
B. A presumptive diagnosis of cyanide poisoning was made.
The symptoms tend to be non-specific, and blood cyanide is
not easy to measure. The almond odor is however
characteristic of gaseous cyanide. Later confirmed that he
has taken a massive dose of amygdalin, obtained from
almonds and containing a cyanide derivative.
C. Treatment: Nitrites, followed by infusion of thiosulfate,
100% oxygen, and sodium bicarbonate. The patient
recovered.
16. Discussion. Cyanide binds to the heme of cytochrome
oxidase, inhibiting the enzyme and blocking respiration.
Nitrites induce the synthesis of methemoglobin and
increase serum levels. Cyanide will also bind to
methemoglobin decreasing the levels available to react
with cytochrome oxidase. Thiosulfate combines with
cyanide to produce thiocyanate which does not react
with the free oxidase. This reaction is mediated by the
mitochondrial enzyme rhodanese. Using this rationale,
cyanide poisoning, while potentially fatal, can be
successfully treated if diagnosed early.
Cyanide PoisoningCyanide Poisoning