20.3 How Is the Electron-Transport Chain Organized?
601
(a) 2 H+ (b) 2 H+ 8/10 7/11/14 13 12
2 e– 2 Fe-S 2 e– UQH2 centers UQ 2 H+
2 Fe-S 2 H+ centers
2 H+ FMNH2 FMN
NAD+ NADH+ + H+
(c) N2
N6b
N6a
N5 ACTIVE FIGURE 20.6 (a) Structural organization of mammalian Complex I, based on electron microscopy, showing functional relationships within the L-shaped complex. Electron flow from NADH to UQH2 in the membrane pool is indicated. (b) Structure of the hydrophilic domain of Complex I from Thermus N1b thermophilus is shown on a model of the membrane-associated complex (pdb id " 2FUG). The locations of N4 N7 individual subunits are indicated. (c) Arrangement of the redox centers in Complex I. The various iron–sulfur N3 centers of Complex I are designated by capital N. (Part a adapted from Janssen, R. J., Nijtmans, L. G., van den Heuvel, L. P., and Smeitink, J. A., 2006. Mitochondrial complex I: Structure, function, and pathology. Journal of Inherited Metabolic Diseases N1a 29:499–515; and parts b and c adapted from Figure 1 of Sazanov, L., and Hinchliffe, P., 2006. Structure of the hydrophilic FMN domain of respiratory Complex I from Thermus thermophilus. Science 311:1430–1436.) Test yourself on the concepts in this figure at www.cengage.com/login.
transport of protons from the matrix side to the cytosolic side of the inner mem- brane. The cytosolic side, where H! accumulates, is referred to as the P (for posi- tive) face; similarly, the matrix side is the N (for negative) face. Some of the energy liberated by the flow of electrons through this complex is used in a coupled process to drive the transport of protons across the membrane. (This is an example of ac- tive transport, a phenomenon examined in detail in Chapter 9.) Available experi- mental evidence suggests a stoichiometry of four H! transported per two electrons passed from NADH to UQ.
Complex II Oxidizes Succinate and Reduces Coenzyme Q
Complex II is perhaps better known by its other name—succinate dehydrogenase, the only TCA cycle enzyme that is an integral membrane protein in the inner mitochondrial membrane. This complex (Figure 20.7) has a mass of 124 kD and is composed of two hydrophilic subunits, a flavoprotein (Fp, 68 kD) and an iron–sulfur protein (Ip, 29 kD), and two hydrophobic, membrane-anchored sub- units (15 kD and 11 kD), which contain one heme b and provide the binding site for UQ. Fp contains an FAD covalently bound to a His residue (see Figure 19.12), and Ip contains three Fe-S centers: a 4Fe-4S cluster, a 3Fe-4S cluster, and a 2Fe-2S cluster. When succinate is converted to fumarate in the TCA cycle, concomitant re- duction of bound FAD to FADH2 occurs in succinate dehydrogenase. This FADH2 602 Chapter 20 Electron Transport and Oxidative Phosphorylation
(a) Intermembrane space Complex II Complex III UQH2 2Fe3+ UQ Heme b
3Fe4S 4Fe4S 2Fe2+ 2Fe2S Matrix 2 H+
FAD FADH2
Succinate Fumarate
(b) (c)
Heme b
Fe-S centers
ACTIVE FIGURE 20.7 (a) A scheme for
electron flow in Complex II. Oxidation of succinate occurs with reduction of [FAD]. Electrons are then passed to Fe-S centers and then to coenzyme Q (UQ). Proton transport does not occur in this complex. (b) The FAD structure of Complex II from pig heart (pdb id " 1ZOY). (c) The arrangement of redox centers. Electron flow is from bottom to top. Test yourself on the concepts in this figure at www.cengage.com/login.
transfers its electrons immediately to Fe-S centers, which pass them on to UQ. Elec- tron flow from succinate to UQ , Succinate ⎯⎯→ fumarate ! 2 H! ! 2 e# (20.18)
yields a net reduction potential of 0.029 V. (Note that the first half-reaction is writ- ten in the direction of the e# flow. As always, !!o$ is calculated according to Equa- tion 20.8.) The small free energy change of this reaction does not contribute to the transport of protons across the inner mitochondrial membrane. This is a crucial point because (as we will see) proton transport is coupled with ATP synthesis. Oxidation of one FADH2 in the electron-transport chain results in synthesis of approximately two molecules of ATP, compared with the approximately three ATPs produced by the oxidation of one NADH. Other flavoproteins can also supply 20.3 How Is the Electron-Transport Chain Organized? 603
C H3C SCoA [FAD]
UQ [FADH2] O
C H3C SCoA H3C CH CH2
FIGURE 20.8 The fatty acyl-CoA dehydrogenase reaction, emphasizing that the reaction involves reduction of CH2 CH CH3 enzyme-bound FAD (indicated by brackets). N
N Fe N
electrons to UQ, including mitochondrial sn -glycerophosphate dehydrogenase, an N _
H3C CH2CH2COO inner membrane-bound shuttle enzyme, and the fatty acyl-CoA dehydrogenases, three soluble matrix enzymes involved in fatty acid oxidation (Figure 20.8; also see _ Chapter 23). The path of electrons from succinate to UQ is shown in Figure 20.7. H3C CH2CH2COO
Iron protoporphyrin IX (found in cytochrome b, Complex III Mediates Electron Transport from Coenzyme Q myoglobin, and hemoglobin) to Cytochrome c In the third complex of the electron-transport chain, reduced coenzyme Q (UQH2) S passes its electrons to cytochrome c via a unique redox pathway known as the Q cycle. UQ–cytochrome c reductase (UQ–cyt c reductase), as this complex is known, involves H3C CHCH3 S three different cytochromes and an Fe-S protein. In the cytochromes of these and sim- ilar complexes, the iron atom at the center of the porphyrin ring cycles between the CH3CH CH3 N reduced Fe2! (ferrous) and oxidized Fe 3! (ferric) states. Cytochromes were first named and classified on the basis of their absorption spec- N Fe N tra (Figure 20.9), which depend upon the structure and environment of their heme groups. The b cytochromes contain iron protoporphyrin IX (Figure 20.10), the same N _ H3C CH2CH2COO heme found in hemoglobin and myoglobin. The c cytochromes contain heme c, derived from iron protoporphyrin IX by the covalent attachment to cysteine residues from the _ H3C CH2CH2COO associated protein. (One other heme variation, heme a, contains a 15-carbon iso- prenoid chain on a modified vinyl group and a formyl group in place of one of the Heme c (found in cytochrome c) methyls [see Figure 20.10]. Cytochrome a is found in two forms in Complex IV of the electron-transport chain, as we shall see.) UQ–cyt c reductase (Figure 20.11) contains a b -type cytochrome, of 30 to 40 kD, with two different heme sites and one c -type cyto- chrome. The two hemes on the b cytochrome polypeptide in UQ–cyt c reductase are distinguished by their reduction potentials and the wavelength ("max) of the so-called
α H3C CH CH2 OH β CH2CH CH3 N (a) N Fe N (a) Cytochrome c: reduced spectrum (b) N _ H3C CH2CH2COO Absorbance
(b) Cytochrome c: oxidized spectrum _
O CH CH2CH2COO
Heme a 450 500 550 600 650 (found in cytochrome a) Wavelength (nm) FIGURE 20.10 The structures of iron protoporphyrin IX, FIGURE 20.9 Visible absorption spectra of cytochrome c. heme c, and heme a.
