Protein engineering cytochrome

Many enzymes have been the subject of protein engineering studies, including several that are important in medicine and industry, eg, lysozyme, trypsin, and cytochrome P450. SubtiHsin, a bacterial serine protease used in detergents, foods, and the manufacture of leather goods, has been particularly well studied (68). This emphasis is in part owing to the wealth of stmctural and mechanistic information that is available for this enzyme. 

Mauk AG (1991) Electron Transfer in Genetically Engineered Proteins. The Cytochrome c Paradigm.75 131-158... 

Wolfbeis OS, Reisfeld R, Oehme 1 (1995) Sol-Gels and Chemical Sensors. 85 51-98 Wong L-L, Westlake ACG, Nickerson DP (1997) Protein Engineering of Cytochrome P450,am. 88 175-208... 

Harford-Cross CF, Carmichael AB, Allan FK et al (2000) Protein engineering of cytochrome P450cam (CYP101) for the oxidation of polycyclic aromatic hydrocarbons. Protein Eng 13 121-128... 

Valderrama B, Garcfa-Arellano H, Giansanti S et al (2006) Oxidative stabilization of iso-1-cytochrome c by redox-inspired protein engineering. FASEB J 20 1233-1235... 

Zvelebil, M. J. J. M., Wolf, C. R., and Sternberg, M. J. E. A predicted three-dimensional structure of human cytochrome P450 implications for substrate specificity. Protein Engineering 4, 271-282 (1991). 

It is quite clear that protein engineering will contribute substantially to future investigations of electron transfer in flavocytochrome 62. To date protein engineering has been used to generate a number of single amino acid substitutions and has allowed the independent expression of the two functionally distinct domains of the enzyme. These two approaches can be readily combined, for example, to express the flavodehydrogenase domain with an active site mutation, thereby sim-plyfing analysis of electron transfer to FMN without interference from the cytochrome domain. 

Because direct electrochemistry is observed only after the problems of interfacial specificity, compatibility, and denaturation have been overcome, it should provide us with a most powerful tool for investigating protein adsorption at surfaces. The studies of genetically engineered cytochrome c variants described in Section II serve as an illustration of this application. 

Hamachi, L, Fujita, A., and Kunitake, T. 1997. Protein engineering using molecular assembly Functional conversion of cytochrome c via noncovalent interactions. Journal ofthe American Chemical Society 119, 9096-9102. 

Erman, J.E. and L.B. Vitello (2002). Yeast cytochrome c peroxidase Mechanistic studies via protein engineering. Biochim. Biophys. Acta 1597, 193-220. 

Loida, P.J. and S.G. Sligar (1993). Engineering cytochrome P-450cam to increase the stereospecificity and coupling of aliphatic hydroxylation. Protein Eng. 6, 207-212. 

Atkins, W.M. and S.G. Sligar (1989). Molecular recognition in cytochrome P450— alteration of regioselective alkane hydroxylation via protein engineering. J. Am. Chem. Soc. Ill, 2715-2717. 

Atkins WM, Sligar SG (1989) Molecular recognition in cytochrome P-450 alteration of regioselec-tive alkane hydroxylation via protein engineering. J Am Chem Soc 111 2715-2717... 

---