This research will establish precisely defined protein structural systems through the application of X-ray crystallographic techniques for the purpose of providing a sound physical basis for biochemical experimentation. The analysis of several proteins to high resolution will be undertaken which initially will include creatine kinase, abrin, catalase, ferritin, canavalin, concanavalin B, and alpha-amylase. Isolation, crystallization, some biochemical characterization and preliminary X-ray diffraction investigations have already been conducted on each of these. Emphasis will be placed on those proteins of most relevant biological interest (creatine kinase, abrin, alpha-amylase, catalase and ferritin) with the objective of elucidating their mechanistic features by examining their structures in the presence of their respective substrates, coenzymes, transition state analogues and effectors by employing the difference Fourier technique. In addition, the studies will examine unique structural features such as a glycoprotein, a virus model, and several unique quaternary structures. A method will be developed for extracting low resolution phase information from electron micrographs for the purpose of expediting X-ray structural analyses. In addition, a system will be set up for digital and optical filtering of electron micrographs.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM027838-07
Application #
3275070
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1979-07-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of California Riverside
Department
Type
Earth Sciences/Resources
DUNS #
City
Riverside
State
CA
Country
United States
Zip Code
92521
Williams, R L; Greene, S M; McPherson, A (1987) The crystal structure of ribonuclease B at 2.5-A resolution. J Biol Chem 262:16020-31
McGrath, M; Cascio, D; Williams, R et al. (1987) Propidium binding to a ribonuclease-DNA complex: X-ray and fluorescence studies. Mol Pharmacol 32:600-5
Axelrod, H; Greene, S M; McPherson, A (1986) X-ray studies of nucleotide binding and pyridoxal phosphate labeling of the gene 5 DNA unwinding protein. Biochim Biophys Acta 870:443-53
McPherson, A; Koszelak, S; Axelrod, H et al. (1986) An experiment regarding crystallization of soluble proteins in the presence of beta-octyl glucoside. J Biol Chem 261:1969-75
McPherson, A; Brayer, G; Morrison, R (1986) Structure of the crystalline complex between ribonuclease A and D(pA)4. Biophys J 49:209-19
McPherson, A; Brayer, G D; Morrison, R D (1986) Crystal structure of RNase A complexed with d(pA)4. J Mol Biol 189:305-27