The recognition and subsequent interactions between a protein and other molecules play essential roles in many fundamental biological processes. The overall objective of this project is to understand the molecular basis of how proteins recognize other molecules, including small molecule ligands as well as macromolecules such as nucleic acids and proteins, using single crystal x-ray diffraction studies. The rationale of the work here is that proteins may possess certain characteristic folding """"""""motifs"""""""" for their substrate binding functions. Specifically, we propose to crystallize and determine the three dimensional structure of several proteins systems. These include 1) proteins that interact with DNA: E. coli Hu protein, Ike and fd gene 5 protein and E. coli Rep enzyme and their complexes with DNA oligonucleotides; 2) Aminoacyl tRNA synthetase fragments and their complexes with tRNA; 3) Fab fragments of antiarsonate immunoglobulins; 4) Synthetic peptides possessing DNA binding activity. We hope to crystallize these proteins with and without their respective substrates so that we can visualize the structure of the protein before and after complex formation. Detailed knowledge of the protein structure will allow us to assess the conformational changes in the protein molecule and to predict ways of altering existing proteins or designing new proteins of various functions. The proposed studies in this project involve collaboration with other principal investigators in the program project. By combining the knowledge of recombinant DNA methodology, monoclonal antibody technology, nucleotide and peptide chemistry and x-ray crystallography within the program project, we hope ultimately to achieve the long range goal of understanding the structure, interactions and function of proteins and of designing new proteins of improved or desired functions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
1P01GM037641-01
Application #
3941959
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Type
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Martinis, S A; Schimmel, P (1992) Enzymatic aminoacylation of sequence-specific RNA minihelices and hybrid duplexes with methionine. Proc Natl Acad Sci U S A 89:65-9
Miller, W T; Schimmel, P (1992) A retroviral-like metal binding motif in an aminoacyl-tRNA synthetase is important for tRNA recognition. Proc Natl Acad Sci U S A 89:2032-5
Miller, W T; Hill, K A; Schimmel, P (1991) Evidence for a ""cysteine-histidine box"" metal-binding site in an Escherichia coli aminoacyl-tRNA synthetase. Biochemistry 30:6970-6
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Musier-Forsyth, K; Scaringe, S; Usman, N et al. (1991) Enzymatic aminoacylation of single-stranded RNA with an RNA cofactor. Proc Natl Acad Sci U S A 88:209-13
Park, S J; Miller, W T; Schimmel, P (1990) Synthetic peptide model of an essential region of an aminoacyl-tRNA synthetase. Biochemistry 29:9212-8
Toth, M J; Schimmel, P (1990) Deletions in the large (beta) subunit of a hetero-oligomeric aminoacyl-tRNA synthetase. J Biol Chem 265:1000-4
Toth, M J; Schimmel, P (1990) A mutation in the small (alpha) subunit of glycyl-tRNA synthetase affects amino acid activation and subunit association parameters. J Biol Chem 265:1005-9
Lai, M Z; Jang, Y J; Chen, L K et al. (1990) Restricted V-(D)-J junctional regions in the T cell response to lambda-repressor. Identification of residues critical for antigen recognition. J Immunol 144:4851-6

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