Molecular chaperones are involved in a wide range of essential cellular processes: protein synthesis, molecular assembly, translocation, degradation, and folding. The E. coli molecular chaperone GroEL, along with its co-chaperone GroES, increases the efficiency of protein folding in vivo, using an ATP-driven mechanism. In the GroEL-facilitated folding process, first GroEL sequesters the aggregation-prone nonnative forms of proteins from the complex cellular environment within its central cavity. Then with the actions of ATP binding/hydrolysis and GroES binding, the protein is allowed to carry out its initial folding events within an isolated """"""""folding chamber"""""""" formed by GroEL/GroES. The long-term goals of this proposal are to elucidate the structural features of the interaction of GroEL and substrate proteins, to understand the mechanism of GroEL-facilitated protein folding in a structural context, and to further structural knowledge of molecular chaperones function in general. Various biochemical and biophysical techniques, including phage display, fluorescence spectroscopy/polarization and X-ray crystallography, will be used to determine how GroEL recognizes the substrate proteins, and to understand the energetics of ATP binding/hydrolysis.
Three specific aims are to: 1) Select for small peptides that interact with the substrate binding domain of GroEL using a phage display method. 2) Study the interplay of GroEL-substrate by determining the structures of the substrate-trapped GroEL assemblies using X-ray protein crystallography and NMR, and carry out structure-guided mutational studies. 3) Create model polypeptide substrates for GroEL based on the peptides selected in 1) and use them to study the functional role of nucleotide binding/hydrolysis and the mechanism of GroEL-assisted protein folding. Knowledge of protein folding and the role of molecular chaperones in facilitating the folding process will contribute to a better understanding of folding-related human diseases, such as Cystic Fibrosis, Alzheimer's, Prion diseases and cataracts, at the molecular level, and could lead to the design of novel therapeutic approaches.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM065260-05
Application #
7222693
Study Section
Biophysical Chemistry Study Section (BBCB)
Program Officer
Smith, Ward
Project Start
2003-05-01
Project End
2010-04-30
Budget Start
2007-05-01
Budget End
2010-04-30
Support Year
5
Fiscal Year
2007
Total Cost
$279,088
Indirect Cost
Name
Indiana University Bloomington
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401
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