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BIOENERGETICS

Yulia Suciati

Krebs Cycle, Electron


Transport and Oxidative
Phosphorylation
Yulia Suciati

SIKLUS KREBS

SIKLUS ASAM SITRAT


Terjadi didalam matriks mitokondria
Proses ini bersifat aerobik
Fungsi utama siklus asam sitrat (siklus
krebs) a/ bekerja sbg lintasan akhir
bersama untuk oksidasi KH, Lipid,
Protein.
Glukosa, as. Lemak, AA, dimetab. Mjd
asetil KoA atau senyawa antara di SAS.

SIKLUS ASAM SITRAT


Step 1: Condensation
In step 1 of the Krebs cycle, the twocarbon compound, acetyl-S-CoA,
participates in a condensation
reaction with the four-carbon
compound, oxaloacetate, to produce
citrate:

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Step 2. Isomerization of Citrate
step 2 involves moving the hydroxyl
group in the citrate molecule so that
we can later form an a-keto acid

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Step 3: Generation of CO2 by an
NAD+ linked enzyme
The Krebs cycle contains two
oxidative decarboxylation steps; this
is the first one
The reaction is catalyzed by the
enzyme Isocitrate dehydrogenase

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Step 4: A Second Oxidative
Decarboxylation Step
This step is performed by a multienzyme complex, the a-Ketoglutarate
Dehydrogenation Complex

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Step 5: Substrate-Level
Phosphorylation

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Step 6: Flavin-Dependent
Dehydrogenation

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Step 7: Hydration of a Carbon-Carbon
Double Bond

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Step 8: A Dehydrogenation Reaction
that will Regenerate Oxaloacetate

HASIL AKHIR S.A.S


12 molekul ATP terbentuk pada
setiap kali putaran S.A.S
Sejumlah ekuivalen pereduksi akan
dialihkan kpd rantai pernafasan dlm
membran dalam mitokondria.

VITAMIN YG PENTING PD S.A.S


Riboflavin, dlm bentuk FAD (Flavin
Adenin Dinukleotida)
Niasin, dlm bentuk NAD
(Nikotinamide Dinukleotida)
Tiamin, dlm bentuk TPP (Tiamin
Pirophosfat)
Asam pantotenat, sbg bag. dr
Koenzim A

FOSFORILASI OKSIDATIF

The figure is found at http://plaza.ufl.edu/tmullins/BCH3023/cell%20respiration.html (December 2006)

Electron Transport Complexes


4 multiprotein complexes in mitochondrial IM

NADH-CoQ (ubiquinone) oxidoreductase


Succinate-CoQ oxidoreductase
Ubiquinone-cytochrome c oxidoreductase
Cytochrome c oxidase - reduction of O 2

Contain a variety of prosthetic groups, iron-sulfur


clusters
Some subunits encoded by mitochondrial DNA

NADH-CoQ (ubiquinone) oxidoreductase


(complex I)
2 electrons passed from NADH,
through FMN, FeS intermediate
electron carriers to ubiquinone
(coenzyme Q)
Ubiquinone - lipid soluble electron
carrier
Proton pumps transport 4 H+ from
matrix to intermembrane space per
pair of electrons
Spatial organization important protons used in reduction of
ubiquinone come from matrix

Succinate-CoQ oxidoreductase (complex II)


Succinate-CoQ oxidoreductase
succinate dehydrogenase is a
component
No protons transported
FAD, FeS serve as intermediate
electron carriers

Ubiquinone-cytochrome c
oxidoreductase (complex III)
Cytochrome c - peripheral protein,
electron carrier
Cytochromes can only accept 1
electron at a time, resulting in Q cycle
2 H+ from 1st Q deposited in
intermembrane space,
1 e- to Cyt c, 1 e- to Qn
2 H+ from 2nd Q deposited
in intermembrane space,
1 e- to Cyt c, 1 e- to Qn
Qn with 2 e- takes 2 H+
from matrix.

Cytochrome c
oxidase
catalyzes reduction of
molecular oxygen
13 subunits
Four protons translocated for
each O2 reduced
Accumulates 4 electrons (Cu+, Fe2+)
for complete reduction before
releasing products or toxic partiallyreduced products
O2 + 4 e- + 4 H+ --> 2 H2O occurs
in matrix, thus removing 4 H+

Chemiosmosis
Chemiosmosis - Movement of protons from high (IMS) to
low
conc (matrix) used to drive ATP synthesis
electrochemical gradient - electrical and chemical
potential
Electron transport drives generation of H+ gradient
+0.14V electrical potential
1.4 pH unit difference
G=~21 kJ/mole H+
H+ gradient drives
ATP synthesis

inner mitochondrial
membrane

ATP synthase

The figure is found at http://plaza.ufl.edu/tmullins/BCH3023/cell%20respiration.html (December 2006)

ATP Synthase

ATP Synthase produces ATP


from ADP & Pi
H+ passage causes conformational
changes (rotation) in F1, leading to
release of ATP so ADP can bind
again
about 3 protons per ATP must pass
through ATP synthase

The Big Picture

small molecule
shuttles
molecules must be transported to and
from matrix
ATP-ADP translocase exports ATP, imports
ADP - movement of more negative ATP
from matrix dissipates electrical potential
across membrane, weakening gradient by
1 H +.
Phosphate translocase uses 1 H+.
cytosolic NADH
DHAP is reduced by NADH to
Glycerol-3-P in muscle
Electrons passed through FAD to Q
is less efficient, but allows transport
against large NADH gradient

malate-aspartate shuttle
malate-aspartate shuttle used in heart, liver,
kidney to transfer cytosolic reducing
equivalents to matrix
No loss in ATP
generation (2.5 ATP
per pair of electrons)

Malate Aspartate Shuttle

http://courses.cm.utexas.edu/emarcotte/ch339k/fall2005/Lecture-Ch19-2/Slide14.JPG

ATP yield/glucose
2 ATP - Glycolysis
3-5 ATP from 2 FADH from 2 NADH from
glycolysis
5 ATP from 2 NADH from transition reaction
15 ATP from 6 NADH from TCA cycle
2 ATP from 2 GTP from TCA cycle
3 ATP from 2 FADH from TCA cycle
30-32 ATP from complete oxidation of
glucose

Inhibitors
Electron flow can be
inhibited by POISONS
Useful in lab to control
entry and exit points for
electron transport studies
Proton gradients are
dissipated by DNP & FCCP,
inhibiting ATP synthesis
Thermogenin in brown
adipose tissue dissipates
proton gradient to
generate heat

Uncoupling
proteins
(UCP)
= separate
RCH from ATP
synthesis
(the synthesis is
interrupted)

energy from H+
gradient is released
as a heat
The figure is found at http://departments.oxy.edu/biology/Franck/Bio222/Lectures/March23_lecture_shuttles.htm (December 2006)

SEMOGA BERMANFAAT
YS 2011

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