Abstract

This project's long-term objective is to broaden and deepen our understanding of enzyme molecules, which are the essential biological catalysts that make life-processes possible. A two-pronged attack is envisioned, combining X-ray crystallography to delineate the 3-dimensional structure of certain target enzymes with site-directed mutagenesis techniques to create mutants in which specified structural modifications have altered the molecule's properties and function. Among the enzymes to be studied are dihydrofolate reductase, cytochrome- c-peroxidase and cytochrome P450(cam).

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM010928-23
Application #
3268181
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1977-01-01
Project End
1987-12-31
Budget Start
1985-01-01
Budget End
1985-12-31
Support Year
23
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of California San Diego
Department
Type
Schools of Arts and Sciences
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093

  Publications

Sawaya, M R; Pelletier, H; Kumar, A et al. (1994) Crystal structure of rat DNA polymerase beta: evidence for a common polymerase mechanism. Science 264:1930-5
Pelletier, H; Sawaya, M R; Kumar, A et al. (1994) Structures of ternary complexes of rat DNA polymerase beta, a DNA template-primer, and ddCTP. Science 264:1891-903
Brown, K A; Howell, E E; Kraut, J (1993) Long-range structural effects in a second-site revertant of a mutant dihydrofolate reductase. Proc Natl Acad Sci U S A 90:11753-6
David, C L; Howell, E E; Farnum, M F et al. (1992) Structure and function of alternative proton-relay mutants of dihydrofolate reductase. Biochemistry 31:9813-22
Farnum, M F; Magde, D; Howell, E E et al. (1991) Analysis of hydride transfer and cofactor fluorescence decay in mutants of dihydrofolate reductase: possible evidence for participation of enzyme molecular motions in catalysis. Biochemistry 30:11567-79
Warren, M S; Brown, K A; Farnum, M F et al. (1991) Investigation of the functional role of tryptophan-22 in Escherichia coli dihydrofolate reductase by site-directed mutagenesis. Biochemistry 30:11092-103
Howell, E E; Booth, C; Farnum, M et al. (1990) A second-site mutation at phenylalanine-137 that increases catalytic efficiency in the mutant aspartate-27----serine Escherichia coli dihydrofolate reductase. Biochemistry 29:8561-9
Howell, E E; Warren, M S; Booth, C L et al. (1987) Construction of an altered proton donation mechanism in Escherichia coli dihydrofolate reductase. Biochemistry 26:8591-8
Villafranca, J E; Howell, E E; Oatley, S J et al. (1987) An engineered disulfide bond in dihydrofolate reductase. Biochemistry 26:2182-9
Howell, E E; Villafranca, J E; Warren, M S et al. (1986) Functional role of aspartic acid-27 in dihydrofolate reductase revealed by mutagenesis. Science 231:1123-8

Showing the most recent 10 out of 12 publications