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Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
5. Oxidative Phosphorylation and
Mitochondria Transport Systems
Mitochondria = power house of the cell
glyco.
TCA NADH, FADH2 (energy rich mols)
f.a.oxi. each has a pair of e- (having a transfer pot.)
2 e-
02 Energy released! (used for ATP)
Oxidative Phoshorylation: the process in which ATP is
formed as electrons are transferred from NADH or FADH2
to O2 by a series of electron carriers
6. Some Features…
1. Oxidative phosphorylation is carried out by respiratory
assemblies that are located in the inner membrane.
– TCA is in the matrix
2. The oxidation of NADH 2.5 ATP
3. FADH2 1.5 ATP
– Oxidation and phosphorylation are COUPLED
4. Respiratory assemblies contain numerous electron
carriers
– Such as cytochromes
5. When electrons are transferred H+ are pumped out
6. ATP is formed when H+ flow back to the mitochondria
7. Some Features Continued…
Thus oxidation and phosphorylation are coupled by
a proton gradient across the inner mitochondria
membrane
– So, we produce ATP through this
– Glycolysis and TCA cycle can continue only if NADH and
FADH2 are somehow reoxidized to NAD+ and FAD
8.
9. Release of Free Energy During Electron
Transport
1. Electrons transferred
electron donor (reductant) electron acceptor (oxidant)
They can be transferred
– H-
– H+
– Pure electrons
2. When a compound loses its electrons becomes
oxidant
cyt b (Fe ++) + cyt c1 (Fe +++) cyt b (Fe +++) + cyt c1 (Fe ++)
red. X oxi. Y oxi. X’ red. Y’
Red. X and Oxi. X’ Redox
Red. Y’ and Oxi. Y Pairs
10. Release of Free Energy Continued…
3. PAIRS differ in their tendency to lose electrons
– It is a characteristic of a pair
– Can be quantitatively specified by a constant… E0 (volts)
– E0: standard reduction potential
– The more negative E0, the higher the tendency of the reductant
to lose electrons
– The more positive E0, the higher the tendency of the oxidant to
accept electrons
– Electron transfer: more –E0 ---------- more +E0
4. Free energy decreases as electrons are transferred
Go = -nF E0
where “n” is the number of electrons transferred, and F is Faraday’s constant (23, 062)
E0 = E0 (electron accepting pair) – E0 (electron donating pair)
11.
12.
13.
14.
15.
16.
17.
18.
19. What Are the Electron Carriers in mt?
Most of the electron carriers in mitochondria are
integral proteins
There are four types of electron transfers
1. Direct transfer of electrons
Fe+3 Fe+2
2. As a hydrogen atom
H+ + electron
3. As a hydride ion
:H- (has 2 electrons)
4. Direct combination of an organic reductant with O2
22. How Is This Order Found?
1. NADH, UQ, cytb, cytc1, c, a, and a3 is the order
– Their standard reduction potentials have been determined
experimentally!
– The order increased E0 because electrons tend to flow from more
negative E0 to more positive E0
2. Isolated mitochondria are incubated with a source of electrons but
without O2
– a, a3 is oxidized first
– c, c1, b are second, third, and fourth respectively
– When the entire chain of carriers is reduced experimentally by
providing an electron source but no O2 (electron acceptor) then O2
suddenly introduced into the system
– The rate at which each electron carrier becomes oxidized shows the
order in which the carriers function
– The carrier nearest O2 is oxidized first, then second, third, etc.
23.
24.
25.
26.
27. Action of Dehydrogenases
Most of the electrons come from
Electron acceptors NAD or FMN, FAD
Reduced subs + NAD+ ox. sub + NADH + H+
Reduced subs + NADP+ ox. Sub + NADPH + H+
In addition to FAD and NAD, there are three other
types of electron carrying groups
– Ubiquinone
– Iron containing proteins (cytochromes, Fe-S proteins)
Ubiquinone = CoQ or = UQ
– When it accepts 1 electron UQH (semiquinone)
– When it accepts 2 electrons UQH2 (ubiquinal)
32. Oxidation states of flavins.
• The reduction of flavin mononucleotide (FMN) to FMNH2
proceeds through a semiquinone intermediate.
33.
34.
35.
36.
37. Complex I
NADH dehydrogenase
(NADH Q reductase)
Huge protein
– 25 pp
FMN, Fe-S
I electron UQ
38.
39. Complex II
Succinate Q Recuctase (Succinate dhydrogenase)
– Is the only membrane bound enzyme in the TCA cycle
– Contains FAD, Fe-S
II electrons UQ
Cytochrome: an electron transferring protein that
contains a heme prosthetic group!
43. Complex III
Cyt reductase (UQ-cyt c oxido reductase or cyt
bc1 complex)
– Contains cyt b, c1, Fe-S proteins and at least six other
protein subunits
UQ is 2e- carrier, cyts are 1e- carriers
– This switch is done in a series of reactions (called Q
cycle)
Electron transfer in III seems to be complicated
but it’s not
Net reaction:
– UQH2 UQ and cyt c is reduced
– H+ is pumped out also
44. Complex IV
Cyto oxidase
– Contains a, a3, and CuA, CuB
The detail of this electron transfer in
Complex IV is not known
It also functions as a proton pump
57. Mitchel’s Theory
The electrochemical potential
difference resulting from the
asymmetric distribution of the H+ is
used to drive the mech. responsible
for the formation of ATP
58.
59.
60.
61. Chemiosmotic Theory Continued…
G = RT ln(C2/C1) + ZF
[ + ] [ + ]
When H+ is pumped against electrochemical gradient
G=+
When protons flow back inside, this G becomes
available to do the work!!
82. Uncoupled Mitochondria in Brown Fat
Produces Heat
This is done by DNP or other uncouplers
They carry protons across the inner mitochondria membrane
In the presence of DNP, electron transport is normal but ATP is
not formed
Proton-motive force is gone or disrupted
Uncoupling is also seen in brown adipose tissue
It is useful to maintain BT in hibernating animals, newborns, and
mammals adapted to the cold
It has lots of mitochondria
IMM thermogenin (uncoupling protein)
Thermogenin generates heat by short-circuiting the mitochondrial
proton battery
83. Shuttle Systems
Required for cytosolic NADH oxidation
NADH dehydrogenase IMM can accept electrons only
from NADH in the matrix
We also make cytosolic NADH by glycolysis
They also have to be reoxidized to NAD+
IMM is not permeable to cytosolic NADH
– We therefore need shuttle systems
Electrons are transferred from NADH to Complex III
(not I), providing only enough energy to make 2 ATP
(G-3-P shuttle)
It is active in muscle (insect flight) and brain
Net reaction:
– NADH + H+ + E- FAD NAD+ + E-FADH2
(cytosolic) (mitochondrial) (cyto) (mito)
So, 2ATP is formed UQ
84.
85.
86.
87. Malate-aspartate Shuttle
Heart
Liver
Cytoplasmic NADH is brought to
mitochondria by this shuttle
This shuttle works only if NADH/NAD+
increase in the cytosol (then
mitochondria)
No energy consumed
No ATP lost
88.
89.
90.
91. Regulation of ATP Producing
Pathways
Coordinately regulated
– Glycolysis
– TCA
– FA oxidation
– a.a. oxidation
– Oxidative phosphorylation
Interlocking regulatory mech.
ATP, ADP controls all of them
Acetyl CoA and and citrate
92.
93.
94. Regulation of Oxidative Phosphorylation
Intracellular [ADP]
If no ADP no ATP
– The dependence of the rate of O2 consumption on the
[ADP] (Pi acceptor) is called “acceptor control”
acceptor control ratio = ADP-induced O2 consumption
O2 consumption without ADP
Mass action ratio:
ATP is high normally
[ADP][Pi]
So, system is fully phosphorylated.
ATP used, ratio decreases, rate of oxidative phosphorylation
increases.
95.
96.
97.
98.
99.
100. Tumor Cells
Regulation is gone in catabolic
processes
Glycolysis is faster than TCA
They use more Glc, but cannot
oxidize pyruvate
Pyruvate lactate
(PH decreases in tm.)
101.
102. Mutations in Mitochondrial Genes
Mutations in mitochondrial genes cause human
disease.
DNA has 37 genes (16, 569 bp), 13 of them encode
respiratory chain proteins.
LHON- Leber’s Hereditary Opti-neuropathy
– CNS problems
– Loss of vision
– Inherited from women.
– A single base change ND4
Arg His (Complex 1)
– Result: defective electron transfer from NADH to UQ.
Succinate UQ okay, but NADH UQ not.
111. Summary
Electron flow results in pumping out H+ and the
generation of membrane potential!
ATP is made when protons flow back to the matrix!
F0F1 complex
Proton motive force, PH gradient, membrane potential
The flow of two electrons through each of three
proton-pumping complexes generates a gradient
sufficient to synthesize one mole of ATP!
112. The proton gradient is an
interconvertible form of free energy
Proton gradients are a central
interconvertible currency of free
energy in biological systems.
• Active transport of Ca
• Rotation of bacterial flagella
• Transfer of e from NADP+ to NADPH
• Generate heat in hybernation