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• W1974506452 abstract "Chemical modification of Escherichia coli phosphoenolpyruvate carboxylase (P-pyruvate carboxylase) by 2,4,6-trinitrobenzene sulfonate, a specific reagent for amino groups, causes desensitization to allosteric inhibitors, l-aspartate and l-malate, as well as inactivation. When l-malate is included in the modification mixture, P-pyruvate carboxylase was markedly protected from both desensitization and inactivation [Naide, A., Izui, K., Yoshinaga, T. & Katsuki, H. (1979) J. Biochem. (Tokyo) 85, 423–432]. To determine the lysine residue(s) involved in allosteric inhibition, the lysine residues that were protected from modification by l-malate were investigated by analyzing trinitrophenylated peptides liberated by digestion with glutamyl endopeptidase (V8-protease). The identified residues were Lys491, Lys620, Lys650, and Lys773. Each of these residues was individually replaced with an alanine or serine residue by site-directed mutagenesis to produce mutant enzymes. The mutant enzyme whose lysine residue was replaced with serine ([Ser620]P-pyruvate carboxylase) showed a marked desensitization to l-aspartate and l-malate, while retaining almost the same maximal catalytic activity as the wild-type P-pyruvate carboxylase. Essentially no changes in enzymatic properties were observed for the [Ala491]- and [Ala650]P-pyruvate carboxylases, while for the [Ala620]- and [Ala773]P-pyruvate carboxylases the polypeptides of the expected size were not significantly accumulated in the transformed E. coli cells, presumably due to intracellular degradation." @default.
• W1974506452 created "2016-06-24" @default.
• W1974506452 creator A5036888298 @default.
• W1974506452 creator A5066679225 @default.
• W1974506452 date "1997-07-01" @default.
• W1974506452 modified "2023-10-17" @default.
• W1974506452 title "The Replacement of Lys620 by Serine Desensitizes Escherichia Coli Phosphoenolpyruvate Carboxylase to the Effects of the Feedback Inhibitors l-aspartate and l-malate" @default.
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• W1974506452 doi "https://doi.org/10.1111/j.1432-1033.1997.t01-1-00074.x" @default.
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