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J. Biol. Chem., Vol. 257, Issue 24, 14603-14605, Dec, 1982
MH O'Leary and E Diaz
Phosphoenol-3-bromopyruvate is an excellent competitive inhibitor (versus
phosphoenolpyruvate) of maize phosphoenolpyruvate carboxylase (Ki = 30
microM), provided that preincubation of enzyme with inhibitor is avoided.
If the enzyme is preincubated with inhibitor in the presence of Mn2+ and
HCO3(-), complete inactivation occurs over the course of about 1 h. The
inactivation is first order in enzyme and is saturable with respect to
inhibitor, with an apparent Michaelis constant of 67.5 microM and a
half-life at high inhibitor concentration of 13.4 min. The inactivation is
inhibited by phospholactate and is much slower at low HCO3(-)
concentration. Incubation of the enzyme with phosphoenol-3-bromopyruvate in
the presence of H14CO3(-) and Mn2+ followed by treatment with NaBH4 leads
to incorporation of 14C into the inactive enzyme. If treatment with NaBH4
is omitted, the enzyme is inactivated, but no 14C is incorporated into the
inactive product. It appears that phosphoenol-3-bromopyruvate is a
mechanism-based inactivator of phosphoenolpyruvate carboxylase, probably
because of enzyme-catalyzed conversion to 3-bromooxalacetate, which
alkylates the enzyme.
Phosphoenol-3-bromopyruvate. A mechanism-based inhibitor of phosphoenolpyruvate carboxylase from maize
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