A second-site mutation at phenylalanine-137 that increases catalytic efficiency in the mutant aspartate-27. fwdarw. serine Escherichia coli dihydrofolate reductase
EE Howell, C Booth, M Farnum, J Kraut… - Biochemistry, 1990 - ACS Publications
EE Howell, C Booth, M Farnum, J Kraut, MS Warren
Biochemistry, 1990•ACS PublicationsRevised Manuscript Received May 22, 1990 abstract: The adaptability of Escherichia coli
dihydrofolate reductase (DHFR) is being explored by identifying second-site mutationsthat
can partially suppress the deleterious effect associated with removal of the active-site proton
donor aspartic acid-27. The Asp27-* serine mutant DHFR (D27S) was previously
characterized and the catalytic activity found to be greatly decreased at pH 7.0 [Howell et
al.(1986) Science 231, 1123-1128], Using resistance to trimethoprim (a DHFR inhibitor) in a …
dihydrofolate reductase (DHFR) is being explored by identifying second-site mutationsthat
can partially suppress the deleterious effect associated with removal of the active-site proton
donor aspartic acid-27. The Asp27-* serine mutant DHFR (D27S) was previously
characterized and the catalytic activity found to be greatly decreased at pH 7.0 [Howell et
al.(1986) Science 231, 1123-1128], Using resistance to trimethoprim (a DHFR inhibitor) in a …
Revised Manuscript Received May 22, 1990 abstract: The adaptability of Escherichia coli dihydrofolate reductase (DHFR) is being explored by identifying second-site mutationsthat can partially suppress the deleterious effect associated with removal of the active-site proton donor aspartic acid-27. The Asp27-* serine mutant DHFR (D27S) was previously characterized and the catalytic activity found to be greatly decreased at pH 7.0 [Howell et al.(1986) Science 231, 1123-1128], Using resistance to trimethoprim (a DHFR inhibitor) in a genetic selection procedure, we have isolated a double-mutant DHFR gene containing Asp27-*•
ACS Publications