The major goal is to understand the structural basis of immune recognition and the detailed chemistry and diversity of antibody- antigen interactions. Two of the most important antibody recognition events which currently require understanding concern antipeptide antibodies and catalytic antibodies. The common theme of these projects is antibodies raised against synthetic antigens, designed as immunological tools to answer important structural questions. The first objective is to determine and refine three- dimensional structures by x-ray crystallography at high resolution (less than 1.9-2.1 A) of an anti-peptide Fab, peptide-Fab complexes and protein-Fab complex in order to understand how an antibody can recognize the same antigenic determinant as a free peptide and as part of an intact viral antigen. The structural recognition of different peptide sequences and specifically designed conformationally-restricted analogues will be correlated with affinity measurements in order to quantitate antibody-antigen interactions. The structures of the free peptide determined by N.M.R. (Dyson et al., 1985), the peptide bound to the Fab and the peptide in the intact influenza virus hemagglutinin (Wilson et al., 1981) will be compared to evaluate antigen conformational changes which occur on antibody binding as well as any changes which occur in the antibody. The second objective is to determine the three dimensional structure by x-ray crystallography of a catalytic Fab and designed transition-state analogue-Fab complexes, and hence, how monoclonal antibodies can act as enzymatic catalysts (abzymes). The catalytic antibody to be studied initially has esterolytic activity and was raised against phosphonate haptens specifically constructed and designed to act as transition state analogues when bound to the antibody. In addition, the structural study will investigate the differential rate enhancement of different esterolytic antibodies. Such results will contribute not only to providing new insights into enzyme catalysis but show how the diversity of the immune system can be manipulated to provide potentially powerful biological, medical and biotechnological tools of exquisite stereochemical specificity. In both these systems crystals of both the free Fab and Fab-antigen complex are available and hence one can evaluate for the first time whether any changes in antibody structure accompany antigen binding.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
7R01AI023498-07
Application #
3135683
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1986-04-01
Project End
1994-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
7
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
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