[BIOCHEMISTRY I] Module No.

2
E
Electron Transport Chain #3.3
Dr. Raymond Oliver A. Cruz September 14, 2016

The Electron transport system contains mainly six components

Outline Dr. Raymond Oliver A. Cruz arranged in the following sequence:
I. SUBSTRATE SHUTTLE
1.NAD (Nicotinamide adenine dinucleotide)
A. Glycerophosphate shuttle Dr. Raymond Oliver A. Cruz 2.FAD (Flavin adenine dinucleotide)
B. Malate shuttle
3.cytochrome B
II. ELECTRON TRANSPORT CHAIN Dr. Raymond Oliver A. Cruz 4.cytochrome C
A. Electron Carriers Dr. Raymond Oliver A. Cruz 5.cytochrome A and
B. Cytochromes
6.cytochrome A3
C. Mitochondria
III. PARTS OF ELECTRON TRANSPORT CHAIN
IV. OXIDATION OF FAD & NADH A. Electron Carriers
V. ATP SYNTHASE • Any of various molecules that are capable of accepting one or
A. ATP structure two electrons from one molecule and donating them to
B. Conformational coupling theory another in the process of electron transport
VI. ATP SYNTHASE • There are 2 sites of entry for electrons into the electron
VII. DRUGS THAT INHIBIT ETC transport chain: NAD or FAD which are coenzymes for
VIII. BROWN ADIPOSE TISSUE dehydrogenase enzymes
IX. DISEASES a.) NAD (Nicotinamide coenzymes)

SUBSTRATE SHUTTLE
• These are the mechanisms by which NADH plus H+ and
FADH2 are used to generate ATP
•
A. Glycerophosphate shuttle
• Mitochondrial glycerol 3- phosphate dehydrogenase is linked
to the respiratory chain via flavoprotein rather than NAD, so
only 1.5 mol rather than 2.5 mol of ATP is formed
• Observed in mammalian MUSCLE and BRAIN
• 2-electron reaction transferring 2 e- and 2 H+
B. Malate shuttle • Derived from vitamin B3 (niacin)
 Found in mammalian KIDNEYS, LIVER and HEART
 More complex but more efficient
 2.5 ATP are formed b.) FAD (The flavin coenzymes / flavoproteins)

II. ELECTRON TRANSPORT CHAIN

• The majority of the energy conserved during catabolism
reactions occurs near the end of the metabolic series of
reactions
• The electron transport or respiratory chain gets its name from
the fact that electrons are transported to meet up with oxygen
from respiration at the end of the chain.
• Transfer of proton (H+) across membrane
If one part stops, the whole process will stop.
• Coupled to ATP formation
• Series of electron carriers
o NADH or FADH2 – as initial donor
o O2 – as final acceptor
• Occurs in the mitochondria, specifically the
inner membrane
•
The overall electron chain transport reaction is:
2 H+ + 2 e+ + 1/2 O2  H2O + energy
• The electron transport chain enzymes are a series of
oxidation-reduction electron carrier molecules and proton
pumps.
• These enzymes use the energy in the electrons from
glycolysis and Krebs cycle to move protons against a
concentration gradient to form the proton motive force.
Proton motive force acts as a proton pump, using the energy of
electrons from an electron carrier (Gibbs free energy of redox reactions)
to pump protons (hydrogen ions) out across the membrane, separating
the charge across the membrane.
• a 2-electron reaction transferring 2 e- and 2 H+
• Derived from vit. B2 (riboflavin)
Coenzyme Q
• (CoQ, Q or ubiquinone) is lipid-soluble. It dissolves in the
hydrocarbon core of a membrane.
• Coenzyme Q is the only electron carrier not bound to a
protein.
B. Cytochromes
• Cytochromes are electron carriers containing hemes.
Hemes in the 3 classes of cytochrome (a, b, c) differ in
substituents on the porphyrin ring.
• Some cytochromes(b,c1,a,a3) are part of large integral
membrane protein complexes.
• Cytochrome c is a small, water-soluble protein.
C. Mitochondria
• The mitochondria, available free energy of respiratory
substrates are captured in great amounts.
Parts of the Mitochondria:
1.) Outer Membrane
• Freely permeable to most ions, small molecules
2.) Inner Membrane :
• Impermeable to most small ions, small and large
molecules
• Allows ions such as OH- , ATP’s ADP’s and other
metabolites to pass through without disrupting the
electrochemical gradient across the membrane.
• ETC mostly occurs at inner membrane
3.) Cristae
• Infoldings of the inner membrane
more infoldings:increased surfacearea

[Guloy, Goco, Gochioco] Checked by: [Gayados] Page 1 of 4

[ETC] [Electron Transport Chain] Module 2, Lecture # 3.3

4.) Matrix : From the cytochromes the electrons are given to the enzyme
• contains enzymes, mRNA, tRNA, mitochondrial ribosomes cytochrome oxidase.

MITOCHONRIAL COMPLEXES Step6:

The cytochrome oxidase finally discharge electron to
oxygen.This oxygen units with hydrogen ions forming water.

Protons (indicated by + charge) enter back into the mitochondrial matrix

through channels in ATP synthase enzyme complex. This entry is
coupled to ATP synthesis from ADP and phosphate (Pi)

Enzymes in the Outer membrane:

• Acyl-CoA synthetase
• Glycerolphosphate acyltransferase
Intermembrane
• Adenylyl kinase
• Creatinine Kinase
Step 7:
Inner Membrane
The cytochrome oxidase finally discharges electron to
• Cardiolipin oxygen. This oxygen units with hydrogen ions forming water
• Enzyme of the Resp. Chain (Complex I-IV)
• ATP Synthase
• Membrane Transporters V. ATP SYNTHASE
A. ATP STRUCTURE
III. PARTS OF ELECTRON TRANSPORT CHAIN

1.) Complex I
• NADH-CoQ Oxidoreductase
• NADH Dehydrogenase
o NADH = initial electron donor
• Gives out 4 H+
2.) Complex II
• SuccinateDehydrogenase
• Succinate–QReductase
o Involved in TCA / Kreb’s
o Components: FAD and 3 Fe-S clusters o Formation of
fumarate from succinate
3.) Complex III
• Cytochromeb/c1complex
• Cytochromecreductase
o Components: cytochrome c, cytochrome b1, cytochrome bh 1. ATP SYNTHASE
& Fe-S Rieske protein • Site of ATP synthesis
4.) Complex IV • ADP + Pi ATP
• CytochromeOxidase • Consist of:
• Cu2+/hemeprotein o F0 subunit – span the membrane and forms the proton
o Components: Heme-containing cytochrome a & cytochrome channel
a3 plus 2 Cu complexes o F1 subunit – made up of several protein subunits which
• Gives out 4 H+ project into the matrix and contain the phosphorylating
5.) Complex V / ATP Synthase mechanism
Produces ATP through the proton gradient
• Utilizes the proton motive force created by the ETC

Please Refer to the Last Page

IV. OXIDATION OF FAD & NADH

Step1:
The initiation of electron transport system is the removal of
hydrogen from the substrate by enzyme NADH
dehydrogenase
2H -> 2H+ + 2e-
the hydrogen atom becomes ionized into protons+ and electrons-

Step2:
The hydrogen ion reduces the co-enzyme NAD
NAD + 2H+ -> NADH +H+

Step3:
NADH is oxidized into NAD by transferring its hydrogen ion to
FAD which act as the hydrogen carrier.

Step4:
From FAD each hydrogen ion is discharged in the cell fluid
and electrons are passed on the cytochromes B,C,A and A3

Step5:

[Guloy, Goco, Gochioco] Checked by: [Gayados] Page 2 of 4

[ETC] [Electron Transport Chain] Module 2, Lecture #

B. CONFORMATIONAL COUPLING THEORY

• Proton gradient leads to conformational changes in the ATP 7. MALIGNANT HYPERTHERMIA

Synthase • Due to inhalation anesthetics like halothane
• Protons are translocated across the membrane, from the • Uncoupling of oxidative phosphorylation from electron
matrix to the intermembrane space transport
• Electrons are transported along the membrane, through a • Decrease ATP, Inc temp, TCA (trichloranisole) stimulated,
series of protein carriers Excess CO2 leading to respiratory acidosis
• Oxygen is the terminal electron acceptor, combining with
electrons and H+ ions to produce water
• As NADH delivers more H+ and electrons into the ETS, the
proton gradient increases, with H+ building up outside the
inner mitochondrial membrane, and OH- inside the
membrane.

VI. DRUGS THAT INHIBIT ETC

1. Amytal rotenone
• it inhibits the transfer of electrons from iron-sulfur centers in
complex I to ubiquinone. This interferes with NADH during the
creation of usable cellular energy (ATP).

2. Antimycin A
• Antimycin A binds to the Qi site of cytochrome c reductase,
thereby inhibiting theoxidation of ubiquinone in the Qi site
thereby disrupting the Q-cycle of enzyme turn over.
Cytochrome c reductase is a central enzyme in the electron
transport chainof oxidative phosphorylation.

3. CN, CO (Cyanide, Carbon Monoxide)

• Carbon Monoxide binds to the iron when it is in the ferrous
state.
• Cyanide is a potent inhibitor at site IV which accounts for
there acute toxicity.

VII. BROWN ADIPOSE TISSUE

• Transfer energy from food into heat

• Involves an uncoupling protein (UCP, also called
thermogenin) which acts as channel for protons
• Both the acute activity of the tissue, the heat production, and
the recruitment process in the tissue (that results in a higher
thermogenic capacity) are under the control of
norepinephrine released from sympathetic nerves.
• Heat production from brown adipose tissue is activated
whenever the organism is in need of extra heat, e.g., post
natally, during entry into a febrile state, and during arousal
from hibernation, and the rate of thermogenesis is centrally
controlled via a pathway initiated in the hypothalamus.
• its thermogenesis enhances neonatal survival and allows for
active life even in cold surroundings.

VIII. DISEASES

1. FATAL INFANTILE MITOCHONDRIAL MYOPATHY

• Decreased activity of mitochondrial DNA-encoded respiratory
complexes
• Progressive liver failure and neurologic abnormalities,
hypoglycemia, increased lactate

2. KEARNS-SAYRE SYNDROME
• Mutations in complex II
• Short stature, ophthalmoplegia, pigmentary retinopathy
• Cardiac conduction defects

3. LEIGHS DISEASE
• mtDNA disorder
• Lactic acidemia, developmental delay, seizure, extraocular
palsies, hypotonia
• Fatal by age 2
• Some with mutations in cytochrome oxidase

4. CYANIDE AND CO
• Combine with cytochrome oxidase

5. 2,4 DINITROPHENOL
• Allows protons from the cytosol to reenter the matrix
• Uncoupled electron transport

6. OLIGOMYCIN
• Binds to the stalk of ATP synthase
• Increases o2 consumption
• Decreased ATP synthesis

[Guloy, Goco, Gochioco] Checked by: [Gayados] Page 3 of 4

[ETC] [Electron Transport Chain] Module 2, Lecture # 3.3

How Electrons are transported Blue Arrow Point of entry for Electrons, (NAD & FAD)
Red Arrow Proteins that accept Electrons & transfers electron to
oxygen (Cyt b, Cyt c1, Cyt c, Cyt a, Cyt a3)
NAD BlueCI Complex I/ NADH Dehydrpgenase
VioletCII Complex II/ Succinate Dehydrogenase
RedCIII Complex III/ CoQ-Cyt C Reducatase
FMN OrangeCIV Complex IV/ Cyt Oxidase
Amytal BlackDrugs that Fe-sIron Sulfur Proteins
rotenone inhibits the
4
Barbiturates complexes
FeS Circle# of protons
trans located

Malonate
Ubiquinone FeS FAD

Antimycin
A 4 Cyt B Cyt C1
According to the ppt. of doc cruz:

NADFMNubiCyt BUbiFeSCyt C1Cyt c Cyt aCyt a31/2 O2

Ubiquinone

CN-CO
Cyt a + Cyt a3 2

½ O2

Quiz:

1. Mobile molecule embedded in the inner mitochondrial

bilipid membrane?
2. Inihibited by Antimycin A?
3. Mobile protein carrier of electrons?
4. Not a proton pump?
5. Has the most negative standard redox potential?
6. Involved in Oxidation of succinate by FAD?
7. Forms ATP?
8. Mutation in Complex II?
9. Name one enzyme found in the inner membrane
10. Step in the Oxidation of FAD & NAD where hydrogen ion
reduces the co-enzyme NAD

10. Step 2
Membrane Transporters
ATP Synthase
IV)
Enzyme of the Resp. Chain (Complex I-
9. Cardiolipin
8. Kearns-Sayre Syndrome
7. Complex V
6. Complex II
5. NADH
4. CII
3. Cyt. C
2. Complex III
1. Ubiquinone

Answers:

[Guloy, Goco, Gochioco] Checked by: [Gayados] Page 4 of 4