Protein Folding and Assembly

Wright, Peter E.

Abstract

This project aims to provide detailed experimental information on the mechanisms and dynamics of protein folding and assembly at the level of peptide fragments. High resolution two-dimensional NMR spectroscopy will be used to investigate the intrinsic folding tendencies of peptide fragments of proteins and to elucidate the factors which lead to their stabilization and assembly into folded protein structures. Previous work has identified incipient secondary structure in peptide fragments derived from influenza virus hemagglutinin (reverse turn) and myohemerythrin (helix). In the present project we will study the influence of amino acid sequence on the stability of the secondary structures in these and other peptides and investigate their folding and stabilization through interactions at the surface of a fully folded carrier protein, bovine pancreatic trypsin inhibitor (BPTI). Each peptide can be coupled at four discrete sites on BPTI giving four different 1:1 peptide: BPTI complexes. This allows us to investigate the influence of several different protein surfaces on the folding of each peptide. NMR techniques will also be used to study the folding and assembly into enzymatically active complexes of peptide fragments of thioredoxin obtained by CNBr and tryptic cleavage.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of General Medical Sciences (NIGMS)
• Type: Research Program Projects (P01)
• Project #: 5P01GM038794-03
• Application #: 3898384
• Support Year: 3
• Fiscal Year: 1989

Institution

• Name: Scripps Research Institute
• City: San Diego
• State: CA
• Country: United States
• Zip Code: 92037

Publications

Karimi, A; Matsumura, M; Wright, P E et al. (1999) Characterization of monomeric and dimeric B domain of Staphylococcal protein A. J Pept Res 54:344-52

Larsen, T A; Olson, A J; Goodsell, D S (1998) Morphology of protein-protein interfaces. Structure 6:421-7

Olson, A J; Pique, M E (1998) Visualizing the future of molecular graphics. SAR QSAR Environ Res 8:233-47

Sikorski, A; Kolinski, A; Skolnick, J (1998) Computer simulations of de novo designed helical proteins. Biophys J 75:92-105

Dyson, H J; Bolinger, L; Feher, V A et al. (1998) Sequence requirements for stabilization of a peptide reverse turn in water solution--proline is not essential for stability. Eur J Biochem 255:462-71

Bai, Y; Karimi, A; Dyson, H J et al. (1997) Absence of a stable intermediate on the folding pathway of protein A. Protein Sci 6:1449-57

Reva, B A; Finkelstein, A V; Sanner, M et al. (1997) Recognition of protein structure on coarse lattices with residue-residue energy functions. Protein Eng 10:1123-30

Sanner, M F; Olson, A J (1997) Real time surface reconstruction for moving molecular fragments. Pac Symp Biocomput :385-96

Reva, B A; Finkelstein, A V; Sanner, M F et al. (1997) Residue-residue mean-force potentials for protein structure recognition. Protein Eng 10:865-76

Kriwacki, R W; Wu, J; Tennant, L et al. (1997) Probing protein structure using biochemical and biophysical methods. Proteolysis, matrix-assisted laser desorption/ionization mass spectrometry, high-performance liquid chromatography and size-exclusion chromatography of p21Waf1/Cip1/Sdi1. J Chromatogr A 777:23-30