Life as aerobes: are there simple rules for activation of dioxygen by enzymes?
Numerous biological systems involve reaction with dioxygen in the absence of readily accessible spectroscopic signals. We have begun to develop a set of "generic" strategies that will allow us to probe the mechanisms of dioxygen activation. In particular, we wish to understand the nature of the dioxygen binding step, the degree to which electron transfer to dioxygen is rate limiting, whether reactive species accumulate during turnover and, finally, whether proton and electron transfer to dioxygen occur as coupled processes. Our strategy will be introduced for an enzyme system that uses only an organic cofactor in dioxygen activation (glucose oxidase). Two key features emerge from studies of glucose oxidase: (1) that formation of the superoxide anion is a major rate-limiting step and (2) that electrostatic stabilization of the superoxide anion plays a key role in catalysis. Similar themes emerge when our protocols are applied to enzymes containing both an active site metal center and an organic cofactor. Finally, enzymes that rely solely on metal centers for substrate functionalization will be discussed. In no instance, thus far, has evidence been found for a direct coupling of proton to electron transfer in the reductive activation of dioxygen.
Klinman
J P
JP
Department of Chemistry, University of California, Berkeley 94720, USA. klinman@socrates.berkeley.edu
D000332:Aerobiosis; D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008084:Lipoxygenase / Q000737:chemistry / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000737:chemistry / Q000378:metabolism*
D005949:Glucose Oxidase; D008084:Lipoxygenase; D014446:Tyrosine 3-Monooxygenase; D006631:Amine Oxidase (Copper-Containing); D010100:Oxygen