The role of glutamate-199 in the aging of cholinesterase.
Aging of organophosphate-conjugated acetylcholinesterase results from the loss of an alkoxy group with concomitant stabilization of the conjugate to spontaneous or nucleophile-induced deacylation. We have examined the kinetics of aging in a pinacolylmethylphosphonofluoridate (soman)-inhibited mutant enzyme in which the glutamate (E199) located at the amino-terminal to the active-site serine (S200) was converted to glutamine (Q). For wild type enzyme, the soman-acetylcholinesterase conjugate aged immediately, giving rise to a form of enzyme resistant to reactivation by oximes. In contrast, the E199Q mutant enzyme was largely resistant to aging and could be reactivated by oximes. Since the pH dependence for aging was not altered appreciably, the primary influence of the loss of charge appears to be on the intrinsic rate of aging. The negative charge on E199 likely imparts an inductive effect on the conjugated organophosphate to facilitate removal of the alkoxy group.
Saxena
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A
Division of Biochemistry, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Doctor
B P
BP
Maxwell
D M
DM
Lenz
D E
DE
Radic
Z
Z
Taylor
P
P
D000110:Acetylcholinesterase / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D002801:Cholinesterase Reactivators / Q000494:pharmacology; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009154:Mutation; D010091:Oximes / Q000494:pharmacology; D011726:Pyridinium Compounds / Q000494:pharmacology; D012999:Soman / Q000494:pharmacology*; D017186:Titrimetry; D014101:Torpedo
D002801:Cholinesterase Reactivators; D005971:Glutamates; D010091:Oximes; D011726:Pyridinium Compounds; D018698:Glutamic Acid; D012999:Soman; D000110:Acetylcholinesterase; C022870:asoxime chloride