Specificity of aspartokinase III from Escherichia coli and an examination of important catalytic residues.
Aspartokinase III (AK III) has been purified from a plasmid-containing strain of Escherichia coli. The enzyme shows broad specificity for the phosphoryl acceptor substrate. Structural analogs of aspartic acid with a derivatized alpha-carboxyl group are accepted as alternative substrates by the enzyme. Derivatives at the alpha-amino group are also tolerated by AK III but with diminished catalytic activity. As has been previously observed with aspartokinase I (T. S. Angeles and R. E. Viola, 1992, Biochemistry 31, 799), derivatization of the beta-carboxyl group, which serves as the phosphoryl acceptor, does not prevent catalytic activity. These beta-derivatized analogs are capable of productive binding to these enzymes through a reversal of regiospecificity, making the alpha-carboxyl group available as the phosphoryl acceptor. Chemical modification and pH profile studies have identified the functional groups of cysteine and histidine as being involved in the catalytic activity of AK III.
Department of Chemistry, University of Akron, Ohio 44325-3601, USA.
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Keng
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Viola
D001222:Aspartate Kinase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003545:Cysteine / Q000032:analysis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013774:Tetranitromethane / Q000494:pharmacology
D004791:Enzyme Inhibitors; D007093:Imidazoles; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D001222:Aspartate Kinase; D013774:Tetranitromethane; D003545:Cysteine; D005033:Ethylmaleimide; C012160:N-acetylimidazole