SITE DIRECTED MUTAGENESIS - A TOOL FOR ENZYME MECHANISM DISSECTION

被引:31
作者
WAGNER, CR
BENKOVIC, SJ
机构
[1] C.R. Wagner and S.J. Benkovic are, the Pennsylvania State University, Department of Chemistry, University Park
关键词
D O I
10.1016/0167-7799(90)90189-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Protein engineering has become the principle means of examining the active site of an enzyme to identify and quantify the roles of specific residues in ligand binding, specificity and catalysis. Site-specific mutagenesis has extended our knowledge gained from X-ray crystallography, and has provided striking proof that the intricate active-site geometry is supported by the remainder of the protein infrastructure for maximum catalytic efficiency. © 1990.
引用
收藏
页码:263 / 270
页数:8
相关论文
共 41 条
[11]   AN INVESTIGATION OF THE CONTRIBUTION MADE BY THE CARBOXYLATE GROUP OF AN ACTIVE-SITE HISTIDINE ASPARTATE COUPLE TO BINDING AND CATALYSIS IN LACTATE-DEHYDROGENASE [J].
CLARKE, AR ;
WILKS, HM ;
BARSTOW, DA ;
ATKINSON, T ;
CHIA, WN ;
HOLBROOK, JJ .
BIOCHEMISTRY, 1988, 27 (05) :1617-1622
[12]   SITE-DIRECTED MUTAGENESIS REVEALS ROLE OF MOBILE ARGININE RESIDUE IN LACTATE-DEHYDROGENASE CATALYSIS [J].
CLARKE, AR ;
WIGLEY, DB ;
CHIA, WN ;
BARSTOW, D ;
ATKINSON, T ;
HOLBROOK, JJ .
NATURE, 1986, 324 (6098) :699-702
[13]  
FALZONE CF, IN PRESS BIOCHEMISTR
[14]   STRUCTURE-ACTIVITY-RELATIONSHIPS IN ENGINEERED PROTEINS - ANALYSIS OF USE OF BINDING-ENERGY BY LINEAR FREE-ENERGY RELATIONSHIPS [J].
FERSHT, AR ;
LEATHERBARROW, RJ ;
WELLS, TNC .
BIOCHEMISTRY, 1987, 26 (19) :6030-6038
[15]   PROBING THE FUNCTIONAL-ROLE OF THREONINE-113 OF ESCHERICHIA-COLI DIHYDROFOLATE-REDUCTASE FOR ITS EFFECT ON TURNOVER EFFICIENCY, CATALYSIS, AND BINDING [J].
FIERKE, CA ;
BENKOVIC, SJ .
BIOCHEMISTRY, 1989, 28 (02) :478-486
[16]   CONSTRUCTION AND EVALUATION OF THE KINETIC SCHEME ASSOCIATED WITH DIHYDROFOLATE-REDUCTASE FROM ESCHERICHIA-COLI [J].
FIERKE, CA ;
JOHNSON, KA ;
BENKOVIC, SJ .
BIOCHEMISTRY, 1987, 26 (13) :4085-4092
[17]   PROGRESS WITH LAUE DIFFRACTION STUDIES ON PROTEIN AND VIRUS CRYSTALS [J].
HAJDU, J ;
JOHNSON, LN .
BIOCHEMISTRY, 1990, 29 (07) :1669-1678
[18]   THE IMPORTANCE OF ARGININE-171 IN SUBSTRATE BINDING BY BACILLUS-STEAROTHERMOPHILUS LACTATE-DEHYDROGENASE [J].
HART, KW ;
CLARKE, AR ;
WIGLEY, DB ;
CHIA, WN ;
BARSTOW, DA ;
ATKINSON, T ;
HOLBROOK, JJ .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 146 (01) :346-353
[19]  
Holbrook JJ, 1975, ENZYMES A, VXI, P191
[20]   FUNCTIONAL-ROLE OF ASPARTIC ACID-27 IN DIHYDROFOLATE-REDUCTASE REVEALED BY MUTAGENESIS [J].
HOWELL, EE ;
VILLAFRANCA, JE ;
WARREN, MS ;
OATLEY, SJ ;
KRAUT, J .
SCIENCE, 1986, 231 (4742) :1123-1128