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Three-dimensional structure of murine anti-p-azophenylarsonate Fab 36-71. 1. X-ray crystallography, site-directed mutagenesis, and modeling of the complex with hapten

Strong, Roland K. and Campbell, Robert and Rose, David R. and Petsko, Gregory A. and Sharon, Jacqueline and Margolies, Michael N. (1991) Three-dimensional structure of murine anti-p-azophenylarsonate Fab 36-71. 1. X-ray crystallography, site-directed mutagenesis, and modeling of the complex with hapten. Biochemistry, 30 (15). pp. 3739-3748. ISSN 0006-2960. https://resolver.caltech.edu/CaltechAUTHORS:20180209-152413551

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Abstract

The structure of the antigen-binding fragment (Fab) of an anti-p-azophenylarsonate monoclonal antibody, 36-71, bearing a major cross-reactive idiotype of A/J mice has been refined to an R factor of 24.8% at a resolution of 1.85 Å. The previously solved partial structure of this Fab a resolution of 2.9 Å. (Rose et al., 1990) was used as an initial model for refinement against the high-resolution data. The complex with hapten has been modeled by docking the small-molecule crystal structure of phenylarsonic acid into the structure of the native Fab on the basis of a low-resolution electron density map of the complex. In this model, residue Arg-96 in the light chain and residues Asn-35, Trp-47, and Ser-99 in the heavy chain contact the arsonate moiety of the hapten; an additional bond is found between the arsonate group and a tightly bound water molecule. The phenyl moiety of the hapten packs against two tyrosine side chains at positions 50 and 106 in the heavy chain. Residue Arg-96 in the light chain had been implicated as involved in hapten binding on the basis of previous experiments, and indeed, this residue appears to play a crucial role in this model. Experiments employing site-directed mutagenesis directly support this conclusion. The heavy-chain complementarity-determining regions have novel conformations not previously observed in immunoglobulins except for the recently solved anti-p-azophenylarsonate Fab R19.9 (Lascombe et al., 1989).


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1021/bi00229a022DOIArticle
https://pubs.acs.org/doi/10.1021/bi00229a022PublisherArticle
Additional Information:© 1991 American Chemical Society. Published in print 16 April 1991. This work was supported by the US. Public Health Service through NIH Grants CA 24432, HL 19259, and AI 23909. J.S. is a recipient of an ACS scholar award. We thank Rebecca To (NRCC) for growing the crystals used for data collection, Dr. Tom Smith (Purdue) for supplying his programs MacFrodo and MacPrep with which many of the figures were prepared, Drs. Greg Farber (Pennsylvania State University) and Barry Stoddard (University of California, Berkeley) for their support and advice, Sanda Teodorescu-Frumosu (Boston University) and Chio-Ying Kao (Boston University) for assistance with some of the experiments involving the characterization of the site-directed mutants, and Payload Systems, (Cambridge, MA) for supplying the DEC Vaxstation 3200.
Funders:
Funding AgencyGrant Number
NIHCA 24432
NIHHL 19259
NIHAI 23909
American Cancer SocietyUNSPECIFIED
Issue or Number:15
Record Number:CaltechAUTHORS:20180209-152413551
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180209-152413551
Official Citation:Three-dimensional structure of murine anti-p-azophenylarsonate Fab 36-71. 1. X-ray crystallography, site-directed mutagenesis, and modeling of the complex with hapten Roland K. Strong, Robert Campbell, David R. Rose, Gregory A. Petsko, Jacqueline Sharon, and Michael N. Margolies Biochemistry 1991 30 (15), 3739-3748 DOI: 10.1021/bi00229a022
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84773
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:13 Feb 2018 21:38
Last Modified:03 Oct 2019 19:22

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