Abstract

We propose a series of structural and mechanistic studies on members of the 70 kilodalton (kDa) heat-shock-like protein family. Based on sequence data for several of these proteins, protein family. Based on sequence data for several of these proteins, plus delineation of the function of the bovine clathrin uncoating ATPase (HSC70), we propose a working model: all 70 kDa heat shock proteins evolved from a common ancestor, maintaining a highly conserved ATPase domain and developing specialized substrate recognition domains. They all should have similar tertiary structures, and all probably catalyze macromolecular rearrangements of some type using the same fundamental mechanism. We will initiate our studies on the representative of this protein family for which a biochemical function is well-understood, bovine HSC70. We will solve the x-ray crystallographic structure of its 44 kDa ATPase core, and will attempt to crystallize and solve the structure of the intact HSC70. We are cloning and will sequence cDNA for the HSC70 mRNA; we will attempt to construct a recombinant overproducer of the protein. We will pursue protein modification (mutagenesis) experiments on HSC70 to address mechanistic questions of substrate recognition and conformational change. A second representative on which we will pursue similar studies in tandem is the bovine glucose regulated protein.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM039928-03
Application #
3297216
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1988-10-01
Project End
1991-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305

  Publications

Xu, Xiaohua; Wang, Shuying; Hu, Yao-Xiong et al. (2007) The periplasmic bacterial molecular chaperone SurA adapts its structure to bind peptides in different conformations to assert a sequence preference for aromatic residues. J Mol Biol 373:367-81
Bitto, Eduard; McKay, David B (2004) Binding of phage-display-selected peptides to the periplasmic chaperone protein SurA mimics binding of unfolded outer membrane proteins. FEBS Lett 568:94-8
Kwon, Ae-Ran; Trame, Christine B; McKay, David B (2004) Kinetics of protein substrate degradation by HslUV. J Struct Biol 146:141-7
Trame, Christine B; McKay, David B (2003) Structure of the Yersinia enterocolitica molecular-chaperone protein SycE. Acta Crystallogr D Biol Crystallogr 59:389-92
Kwon, Ae-Ran; Kessler, Benedikt M; Overkleeft, Herman S et al. (2003) Structure and reactivity of an asymmetric complex between HslV and I-domain deleted HslU, a prokaryotic homolog of the eukaryotic proteasome. J Mol Biol 330:185-95
Bitto, Eduard; McKay, David B (2003) The periplasmic molecular chaperone protein SurA binds a peptide motif that is characteristic of integral outer membrane proteins. J Biol Chem 278:49316-22
Bitto, Eduard; McKay, David B (2002) Crystallographic structure of SurA, a molecular chaperone that facilitates folding of outer membrane porins. Structure 10:1489-98
Sousa, Marcelo C; Kessler, Benedikt M; Overkleeft, Herman S et al. (2002) Crystal structure of HslUV complexed with a vinyl sulfone inhibitor: corroboration of a proposed mechanism of allosteric activation of HslV by HslU. J Mol Biol 318:779-85
Wedekind, J E; Trame, C B; Dorywalska, M et al. (2001) Refined crystallographic structure of Pseudomonas aeruginosa exotoxin A and its implications for the molecular mechanism of toxicity. J Mol Biol 314:823-37
Sousa, M C; McKay, D B (2001) Structure of Haemophilus influenzae HslV protein at 1.9 A resolution, revealing a cation-binding site near the catalytic site. Acta Crystallogr D Biol Crystallogr 57:1950-4

Showing the most recent 10 out of 34 publications