The goal of the proposed research is to understand the structural basis of recognition of viral antigens and synthetic peptides by the immune system. In particular, the proposal is to determine the three-dimensional structures by x-ray crystallography of an anti-peptide Fab, and Fab- peptide complex and a Fab- protein complex to understand recognition of the same antigenic determinant in a pepetide and in a protein. This nine amino acid determinant (98-106 HA1) has been shown by several antipeptide monoclonal antibodies to be the major immunodominant site of a 36 amino acid synthetic immunogen corresponding to sequence 75-110 of the A/Vic/3/75 strain of influenza virus haemagglutinin (HA1). The conformation of this determinant has been analyzed in the structure of the solved Hong Kong HA structure and in solution by NMR. The main goals are to determine the following structures for a complete description of the antibody-antigen recognition. 1. Crystal structure of antipeptide Fab. The three dimensional structure will be determined from the currently available x-ray data quality crystals of a monoclonal Fab raised against a 36 amino acid synthetic peptide corresponding to residues 75-110 of A/Vic/3/75 haemagglutinin (HA1) of influenza virus. 2. Crystal structure of a Fab- peptide complex. Crystals of the Fab will be soaked in solutions of various synthetic peptides (about 97-108 HA1, Ka 10-6 - 10-8) and the currently available crystals grown from a mixture of the Fab and peptide (88-110 HA1) will be used to determine the Fab- peptide complex structure. 3. Crystal structure of an Fab- protein. Crystallization conditions of mixtures of the anti-peptide Fab and the HA1 """"""""tops"""""""" (HA1 27-328. Ka about 10-6 - 10-7) will be sought to obtain large single crystals of the complex. Structural studies of suitable crystals will be initiated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023498-03
Application #
3135679
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1986-04-01
Project End
1989-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
San Diego
State
CA
Country
United States
Zip Code
92037
Stura, E A; Ghosh, S; Garcia-Junceda, E et al. (1995) Crystallization and preliminary crystallographic data for class I deoxyribose-5-phosphate aldolase from Escherichia coli: an application of reverse screening. Proteins 22:67-72
Haynes, M R; Stura, E A; Hilvert, D et al. (1994) Routes to catalysis: structure of a catalytic antibody and comparison with its natural counterpart. Science 263:646-52
Churchill, M E; Stura, E A; Pinilla, C et al. (1994) Crystal structure of a peptide complex of anti-influenza peptide antibody Fab 26/9. Comparison of two different antibodies bound to the same peptide antigen. J Mol Biol 241:534-56
Haynes, M R; Stura, E A; Hilvert, D et al. (1994) Crystallization and preliminary structural studies of a chorismate mutase catalytic antibody complexed with a transition state analog. Proteins 18:198-200
Wilson, I A; Ghiara, J B; Stanfield, R L (1994) Structure of anti-peptide antibody complexes. Res Immunol 145:73-8
Schulze-Gahmen, U; Rini, J M; Wilson, I A (1993) Detailed analysis of the free and bound conformations of an antibody. X-ray structures of Fab 17/9 and three different Fab-peptide complexes. J Mol Biol 234:1098-118
Kemble, G W; Bodian, D L; Rose, J et al. (1992) Intermonomer disulfide bonds impair the fusion activity of influenza virus hemagglutinin. J Virol 66:4940-50
Rini, J M; Schulze-Gahmen, U; Wilson, I A (1992) Structural evidence for induced fit as a mechanism for antibody-antigen recognition. Science 255:959-65
Wilson, I A; Stanfield, R L; Rini, J M et al. (1991) Structural aspects of antibodies and antibody-antigen complexes. Ciba Found Symp 159:13-28;discussion 28-39
Wilson, I A; Rini, J M; Fremont, D H et al. (1991) X-ray crystallographic analysis of free and antigen-complexed Fab fragments to investigate structural basis of immune recognition. Methods Enzymol 203:153-76

Showing the most recent 10 out of 17 publications