Electron transfer oxygenases

Cytochrome P-450 has been characterized in four stable states [Fe, Fe " RH, Fe RH, (O2—Fe ) RH (metastable)] during its oxygenase reaction cycle. In the complete native system a flavoprotein and a redoxin (putidaredoxin) act as electron donors but also as effectors that complement the cytochrome. In the more complex microsomal system the sequence and intermediates are less well defined the electron-transfer chain contains two flavoproteins and one cytochrome, whose interactions with cytochrome P-450 are still the subject of great controversy. 

NADPH oxidation and NO synthesis by the enzyme, it supports a role for reduction of the heme iron in catalysis, and may explain why NOS functions only as an NADPH-dependent reductase in the absence of bound calmodulin (Klatt et ai, 1993). The mechanism of calmodulin gating is envisioned to involve a conformational change between the reductase and oxygenase domains of NOS, such that an electron transfer between the terminal flavin and heme iron becomes possible. Calmodulin may also have a distinct role within the NOS reductase domain, in that its binding dramatically increases reductase activity of the enzyme toward cytochrome c (Klatt et al., 1993 Heinzel et al., 1992). However, it is clear that several other NOS functions occur independent of calmodulin, including the binding of L-arginine and NADPH, and transfer of NADPH-derived electrons into the flavins (Abu-Soud and Stuehr, 1993). 

The formation of the monodehydroascorbate free radical by dopamine P-mono-oxygenase was similarly demonstrated by the acceleration of the reduction of Fe(III)-cytochrome c and by direct spectrophotometry at 360 nm The action of the laccase from the mushroom Agaricus hisporum on catechol yielded also the semiquinone as the result of a one-electron transfer... 

A large number of investigations of the mechanism of electron transfer reactions of macromolecule-metal complexes in biological systems has been reported. These investigations were concerned with not only natural metalloenzymes such as cytochromes, ferredoxin, blue coppers, oxygenase, peroxidase, catalase, hemoglobin, and ruberodoxin, but also modified metalloenzymes 47). 

A molecular conversion system based on a four-electron transfer to 02 was accomplished in the 02-oxidative polymerization of diphenyl disulfide (Figure 14) [116]. This is the first example of a multielectron mediator that is applied to molecular conversion systems. The multielectron transfer process from the reduced vanadium(III) complex (VOV+) to 02 not only revealed the 02 oxidation mechanism but also provided additional insight into the unique chemistry of vanadium with possible relevance to metal mono-oxygenases. 

The enzymes appear to be cytochrome Ph50 mixed-function oxygenases with electron transfer components similar to those found in liver (19,20). 

The differences in structures and redox partners between the two classes of P450 and NOS enzymes give rise to differences in reduction potentials and electron transfer mechanisms. The fusion of the oxygenase domain to its diflavin reductase domain facilitates ET (of relevance here is that P450 BM3 has the highest mono-oxygenase activity of all P450s [56]). 

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