The long term objectives of the proposed research are related to understanding the role of protein-protein interactions in the regulation of certain metabolic processes and to understanding structure-function relationships in enzymes. Specific projects and 1) the polymerization of actin and the role of actin binding proteins in the polymerization process as well as in the structure and nature of actin gels in both in vitro and in vivo systems; 2) the characterization of tubulin from erytrocytes; 3) the mechanism of action of adenosine deaminase and dihydrofolate reductase; and 4) the kinetics and mechanism of protein folding using wild type and site directed mutants of dihydrofolate reductase. Work with actin, actin binding proteins and tubulin is directly related to various cellular functions in non-muscle cells including cell motility, cytoskeletal structure, endocytosis, cell division and so on. Of particular interest are proposed experiemnts to measure the consequences of known shear stresses on cellular dynamics using digital imaging techniques. Both adenosine deaminase and dihydrofolate reductase are targets for inhibitory therapeutic drugs. Inhibitors of adenosine deaminase have been used in patients with T-cell acute lymphoblastic leukemia while inhibitors of dihydrofolate reductase have been used both as antibacterial agents and anticancer agents presumably because the tetrahydrofolate produced is ultimately related to DNA synthesis. The folding experiments proposed with wild type and site directed mutants of dihydrofolate reductase may lead to a better undrstanding of structure-function relationships in proteins and subsequently to the ability to create new enzymes with specific functions.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37DK013332-24
Application #
3483012
Study Section
Special Emphasis Panel (NSS)
Project Start
1977-01-01
Project End
1994-12-31
Budget Start
1992-02-01
Budget End
1992-12-31
Support Year
24
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Garai, Kanchan; Baban, Berevan; Frieden, Carl (2011) Dissociation of apolipoprotein E oligomers to monomer is required for high-affinity binding to phospholipid vesicles. Biochemistry 50:2550-8
Garai, Kanchan; Frieden, Carl (2010) The association?dissociation behavior of the ApoE proteins: kinetic and equilibrium studies. Biochemistry 49:9533-41
Mustafi, Sourajit M; Garai, Kanchan; Crick, Scott L et al. (2010) Substoichiometric inhibition of Abeta(1-40) aggregation by a tandem Abeta(40-1-Gly8-1-40) peptide. Biochem Biophys Res Commun 397:509-12
Niu, Weiling; Shu, Qin; Chen, Zhiwei et al. (2010) The role of Zn2+ on the structure and stability of murine adenosine deaminase. J Phys Chem B 114:16156-65
Garai, Kanchan; Crick, Scott L; Mustafi, Sourajit M et al. (2009) Expression and purification of amyloid-beta peptides from Escherichia coli. Protein Expr Purif 66:107-12
Zhang, Rui; Hu, Xiaoyan; Khant, Htet et al. (2009) Interprotofilament interactions between Alzheimer's Abeta1-42 peptides in amyloid fibrils revealed by cryoEM. Proc Natl Acad Sci U S A 106:4653-8
Frieden, Carl (2007) Protein aggregation processes: In search of the mechanism. Protein Sci 16:2334-44
Li, Hua; Frieden, Carl (2007) Observation of sequential steps in the folding of intestinal fatty acid binding protein using a slow folding mutant and 19F NMR. Proc Natl Acad Sci U S A 104:11993-8
Li, Hua; Frieden, Carl (2007) Comparison of C40/82A and P27A C40/82A barstar mutants using 19F NMR. Biochemistry 46:4337-47
Chattopadhyay, Krishnananda; Frieden, Carl (2006) Steady-state and time-resolved fluorescence studies of the intestinal fatty acid binding protein. Proteins 63:327-35

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