Cytochrome catalytic cycle

N—Fe(IV)Por complexes. Oxo iron(IV) porphyrin cation radical complexes, [O—Fe(IV)Por ], are important intermediates in oxygen atom transfer reactions. Compound I of the enzymes catalase and peroxidase have this formulation, as does the active intermediate in the catalytic cycle of cytochrome P Q. Similar intermediates are invoked in the extensively investigated hydroxylations and epoxidations of hydrocarbon substrates cataly2ed by iron porphyrins in the presence of such oxidizing agents as iodosylbenzene, NaOCl, peroxides, and air. 

Scheme 10.4 The catalytic cycle of cytochrome P450. Only one possible valence structure of the oxoferrous species IV has been depicted for clarity. See text for details.

Moreover, an electron transfer chain could be reconstituted in vitro that is able to oxidize aldehydes to carboxylic acids with concomitant reduction of protons and net production of dihydrogen (213, 243). The first enzyme in this chain is an aldehyde oxidoreductase (AOR), a homodimer (100 kDa) containing one Mo cofactor (MOD) and two [2Fe—2S] centers per subunit (199). The enzyme catalytic cycle can be regenerated by transferring electrons to flavodoxin, an FMN-con-taining protein of 16 kDa (and afterwards to a multiheme cytochrome and then to hydrogenase) ... 

Michel H. 1999. Cytochrome c oxidase Catalytic cycle and mechanisms of proton pumping— A discussion. Biochemistry 38 15129. 

Figure 2.5 Catalytic cycle of cytochrome P450 including postulated structures of putative intermediates. RH represents the substrate and R(0)H the product. The porphyrin ring is abbreviated as a parallogram with nitrogens at the comers. Adapted with permission from Sato et ah, 1996. Copyright (1996) American Chemical Society.

Figure 9.6. Proposed cytochrome P-450 catalytic cycle (S = cysteine).

It has been proposed [2] (Figure 24.1) that after binding to cytochrome, the substrates such as epoxides, A-oxides, nitro compounds, and lipid hydroperoxides accept two electrons and are reduced to the compounds RH(H)2. In contrast, the oxidizable substrates react with the oxygenated P-450 complex (RH)Fe2+02 (RH)Fe3+ 02 -. After transfer the second electron substrate RH is hydroxylated to ROH and cytochrome P-450 is oxidized to the starting Fe3+ state, completing the catalytic cycle. It is possible that hydroxylation proceeds through the formation of hydroxyl and carbon radicals [3], but a true role of free radicals at the final stages of hydroxylation is still obscure. 

The answers are 34-g, 35-a, 36-d. (Katzung, pp 53—56J There are four major components to the mixed-function oxidase system (1) cytochrome P450, (2) NAD PH, or reduced nicotinamide adenine dinucleotide phosphate, (3) NAD PH—cytochrome P450 reductase, and (4) molecular oxygen. The figure that follows shows the catalytic cycle for the reactions dependent upon cytochrome P450. 

Sulfite oxidase is a dimetallic enzyme that mediates the two-electron oxidation of sulfite by the one-electron reduction of cytochrome c. This reaction is physiologically essential as the terminal step in oxidative degradation of sulfur compounds. The enzyme contains a heme cofactor in the 10 kDa N-terminal domain and a molybdenum center in the 42 kDa C-terminal domain. The catalytic cycle is depicted in Fig. 9. 

The two-electron oxidation of sulfite generates a Molv Fem state, which converts to a Mov-Fen state. Cytochrome c then oxidizes this state to Mov-Fem, which then undergoes another internal electron transfer (k3) to form MoVI-Fen. A second oxidation by cytochrome c forms MoVI-Fem, which completes the catalytic cycle. 

Most published cytochrome P450 catalytic cycles will show the end-on peroxo coordination as shown in Figure 7.14 (4,5a, 5b). Shaik and co-workers have used density functional theory methods to propose a two-state theory... 

S.3 Cytochrome P450 Model Compounds Functional. Ferric-peroxo species are part of the cytochrome P450 catalytic cycle as discussed previously in Section 7.4.4. For instance, these ferric-peroxo moieties are known to act as nucleophiles attacking aldehydic carbon atoms in oxidative deformylations to produce aromatic species.An example of this work, establishing the nucleophilic nature of [(porphyrin)Fe (02)] complexes, was achieved for alkene epoxidation reactions by J. S. Valentine and co-workers. The electron-deficient compound menadione (see Figure 7.18) yielded menadione epoxide when reacted with a [(porphyrin)Fe X02)] complex. 

Substitution of zinc(ll) ions into cytochrome c peroxidase (ZnCcP) has been used to exploit photoactivation of electron transfer (eT) reactions since the mid-1990s. The ZnCcP triplet state ( ZnCcP) reduces Fe(III) cytochrome c, and then back electron transfer recombines the charge separation to complete the catalytic cycle (see Figure 7.36). 

Fig. 5. Catalytic cycle of cytochrome P450. The substrate HR binds to the resting enzyme A to form intermediate B, which is reduced by one electron to form C and then reacts with dioxygen. The resulting ferric-peroxo intermediate D is reduced by one equivalent to form the transient oxyferrous intermediate E, which proceeds quickly to intermediate F with release of a molecule of water. F is designated Fe(V)=0 to indicate that it is oxidized by two equivalents greater than A and not to imply anything about the true oxidation state of the iron. Intermediate F then transfers an oxygen atom to the substrate to regenerate the resting enzyme. The peroxide shunt refers to the reaction of B with hydrogen peroxide to produce the intermediate F, which can then proceed to product formation.

Such an involvement of an amino acid side-chain ligand switch within each catalytic cycle was a novel proposal and as such needs to be scrutinized by a variety of experimental procedures as well as analysis in the context of information known for cytochrome cd nitrite reductase from another source (see later discussion). However, it is interesting to note that something similar has been proposed for the protocate-chuate 3,4-dioxygenase enzyme from Pseudomonas putida (15). On the other hand, bacterial cytochrome c peroxidase offers an example where ligand switching seemingly relates only to an activation phenomenon. 

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