In silico design of a mutant of cytochrome P450 containing selenocysteine.
A mutant of P450(cam), in which the cysteine ligand was replaced by selenocysteine, was designed theoretically using hybrid QM/MM (quantum mechanical/molecular mechanical) calculations. The calculations of the active species, Se-CpdI (selenocysteine-Compound I), of the mutant enzyme indicate that Se-Cpd I will be formed faster than the wild-type species and be consumed more slowly in C-H hydroxylation. As such, our calculations suggest that Se-Cpd I can be observed unlike the elusive species of the wild-type enzyme (Denisov, I. G.; Makris, T. M.; Sligar, S. G.; Schlichting, I. Chem. Rev. 2005, 105, 2253-2277). Spectral features of Se-Cpd I were calculated and may assist such eventual characterization. The observation of Se-Cpd I will resolve the major puzzle in the catalytic cycle of a key enzyme in nature.
Cohen
Shimrit
S
Department of Organic Chemistry, The Hebrew University, Jerusalem, Israel.
Kumar
Devesh
D
Shaik
Sason
S
D019943:Amino Acid Substitution; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006860:Hydrogen Bonding; D006900:Hydroxylation; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D011789:Quantum Theory; D017279:Selenocysteine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics
D017279:Selenocysteine; D003577:Cytochrome P-450 Enzyme System