A Conserved Alcohol Side Chain in the Active Site of

A Conserved Alcohol Side Chain in the Active Site of P450 

Upon examination of aligned sequences of P450 isozymes, it has become clear that an 

Ala245Ser and AIa245Thr mutant enzymes, it appears as though the loss of catalytic activity in the mutant forms correlates with the loss or repositioning of Wat519 (ref [234]). Nonetheless, these mutants are still able to metabolize the substrate testosterone at different sites, implying that a very intricate hydrogenbonding interplay exists between substrate and enz)mie. 

Thr252Ala/Asp251 Asn, in which the stoichiometric ratio of the two pathways is equal at room temj erature . As primary proton transfer is also visible at low temperature in this mutant construct, prior to decay to the resting state, one is able to definitely assess that the protonated peroxo intermediate serves as the branching intermediate of these two pathways. 

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