Abstract

The proposed research is designed to gain insights into the structure function relationships in proteins. We have developed a model system in which the protein under study is a murine immunoglobulin, specific for the hapten, pazobenzenearsonate. We are able to isolate in essentially unlimited numbers and essentially unlimited quantities, mutants of a single antibody that have increased their affinity for the hapten. Similarly, we are able to isolate mutants that have lost their reactivity to antigen. Preliminary results employing x-ray crystallographic methods has demonstrated the feasibility of using this technique for structural analysis in this system. We will also employ the techniques of genetic engineering, site- directed mutagenesis and computer assisted model building to produce variants of the immunoglobulin molecule that will serve to verify deductions concerning structural rules for proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM037641-04
Application #
3877928
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
Massachusetts Institute of Technology
Department
Type
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139

  Publications

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
Kamradt, T; Soloway, P D; Perkins, D L et al. (1991) Pertussis toxin prevents the induction of peripheral T cell anergy and enhances the T cell response to an encephalitogenic peptide of myelin basic protein. J Immunol 147:3296-302
Musier-Forsyth, K; Usman, N; Scaringe, S et al. (1991) Specificity for aminoacylation of an RNA helix: an unpaired, exocyclic amino group in the minor groove. Science 253:784-6
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

Showing the most recent 10 out of 17 publications