An arginine residue at the active site of beta-lactamase from Shigella flexneri UCSF-129.
The role of a residue of arginine at the active site of beta-lactamase from Shigella flexneri UCSF-129 was studied. It is a local pathogenic strain which produces intestinal problems, especially in children. Purified enzymes were obtained by affinity chromatography on phenylboronic acid-agarose gels. The enzyme was serine dependent with a molecular weight of 23.6 kD. It was specifically modified with phenylglyoxal (1/830 molar ratio) and incubated for 20 min in the presence of 50 mM sodium phosphate buffer at pH 8.3. The chemical change was established by isoelectric focusing, since a loss of one positive charge was detected. Protection by cephradine, a substrate, indicated the presence of a residue of arginine at its active site. Controls were conducted by differential spectroscopy. Similar results were obtained with 2,3,butanedione. This vital arginine stabilizes the negative charge of the carboxylic group of C-3 or C-4 from the substrates.
Campos
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Department of Chemistry, Faculty of Sciences, University of Concepción, Chile.
Alarcón
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MA
González
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H
Carvajal
N
N
D001120:Arginine / Q000737:chemistry*; D002515:Cephradine / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D010658:Phenylglyoxal / Q000494:pharmacology; D012763:Shigella flexneri / Q000145:classification / Q000201:enzymology*; D001618:beta-Lactamases / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification
D001120:Arginine; D001618:beta-Lactamases; D002515:Cephradine; D010658:Phenylglyoxal