title: Critical hydrogen bonding by serine 235 for cephalosporinase activity of TEM-1 beta-lactamase.
abstract: The role of Ser-235 in the catalytic mechanism of the TEM-1 beta-lactamase has been explored by the study of a mutant enzyme in which Ser-235 has been substituted by alanine (Ala-235 mutant enzyme). A comparative kinetic analysis of both the wild-type and the Ala-235 TEM-1 enzymes revealed little effect of this substitution of residue 235 on the turnover of penicillins but a greater effect on the turnover of cephalosporins. Susceptibility testing of Escherichia coli strains harboring the wild-type TEM-1 beta-lactamase and the Ala-235 mutant enzyme revealed an effect of the mutation similar to that observed in the enzymological studies. The MICs of two representative cephalosporins for the strain containing the mutant enzyme were much lower than those for the isogenic strain bearing the wild-type TEM-1 beta-lactamase. On the other hand, the strain with the mutant enzyme was still highly resistant to penicillins.
authors.affiliation: Department of Chemistry, Wayne State University, Detroit, Michigan 48202.
authors.forename: U
authors.initials: U
authors.lastname: Imtiaz
authors.forename: E K
authors.initials: EK
authors.lastname: Manavathu
authors.forename: S A
authors.initials: SA
authors.lastname: Lerner
authors.forename: S
authors.initials: S
authors.lastname: Mobashery
mesh_terms: D001483:Base Sequence; D002510:Cephalosporinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002511:Cephalosporins / Q000378:metabolism; D002942:Circular Dichroism; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012694:Serine / Q000737:chemistry*; D001618:beta-Lactamases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D002511:Cephalosporins; D012694:Serine; D002510:Cephalosporinase; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1