FUNCTION OF THE CONSERVED TRIAD RESIDUES IN THE CLASS-C BETA-LACTAMASE FROM CITROBACTER-FREUNDII GN346

被引:27
作者
TSUKAMOTO, K [1 ]
NISHIDA, N [1 ]
TSURUOKA, M [1 ]
SAWAI, T [1 ]
机构
[1] CHIBA UNIV,FAC PHARMACEUT SCI,DIV MICROBIAL CHEM,1-33 YAYOICHO,CHIBA 260,JAPAN
关键词
Active site; Cephalosporinase; Citrobacter freundii; Conserved residue; Site-directed mutagenesis; β-Lactamase;
D O I
10.1016/0014-5793(90)80416-G
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The conserved KTG triad in the class C β-lactamase from Citrobacter freundii GN346 was examined as to its function by means of site-directed mutagenesis. The following conversions were performed; Lys-315 to arginine, alanine or glutamic acid, Thr-316 to valine, and Gly-317 to alanine, proline or isoleucine. The resultant mutant enzymes revealed that a basic amino acid at position 315 and a small uncharged residue at position 317 are essential for the enzyme activity, but a hydroxyl group at residue 316 is not required for the enzymatic catalysis. The kinetic properties of the purified Arg-315 and Val-316 enzymes provided information on the function of these residues. © 1990.
引用
收藏
页码:243 / 246
页数:4
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