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Title: A mutagenesis study of a catalytic antibody

Abstract

The authors have generated seven site-specific mutations in the genes encoding the variable region of the heavy chain domain (V{sub H}) of the phosphocholine-binding antibody S107.S107 is a member of a family of well-characterized highly homologous antibodies that bind phosphorylcholine mono- and diesters. Two of these antibodies, MOPC-167 and T15, have previously been shown to catalyze the hydrolysis of 4-nitrophenyl N-trimethylammonioethyl carbonate. Two conserved heavy-chain residues, Tyr-33 and Arg-52, were postulated to be involved in binding and hydrolysis of 4-nitrophenylcholine carbonate esters. To more precisely define the catalytic roles of these residues, three Arg-52 mutants (R52K, R52Q, R52C) and four Tyr-33 mutants (Y33H, Y33F, Y33E, Y33D) of antibody S107 were generated. The genes encoding the V{sub H} binding domain of S107 were inserted into plasmid pUC-fl, and in vitro mutagenesis was performed. These results not only demonstrate the importance of electrostatic interactions in catalysis by antibody S107 but also show that catalytic side chains can be introduced into antibodies to enhance their catalytic efficiency.

Authors:
; ; ;  [1]
  1. (Univ. of California, Berkeley (United States))
Publication Date:
OSTI Identifier:
5044075
DOE Contract Number:  
AC03-76SF00098
Resource Type:
Journal Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America; (United States)
Additional Journal Information:
Journal Volume: 88:1; Journal ID: ISSN 0027-8424
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ANTIBODIES; MUTAGENESIS; GENE MUTATIONS; DNA SEQUENCING; GENETIC MAPPING; HEAVY WATER; OLIGONUCLEOTIDES; PHOSPHORUS 32; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; DAYS LIVING RADIOISOTOPES; HYDROGEN COMPOUNDS; ISOTOPES; LIGHT NUCLEI; MAPPING; MUTATIONS; NUCLEI; NUCLEIC ACIDS; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; OXYGEN COMPOUNDS; PHOSPHORUS ISOTOPES; RADIOISOTOPES; STRUCTURAL CHEMICAL ANALYSIS; WATER; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Jackson, D.Y., Prudent, J.R., Baldwin, E.P., and Schultz, P.G. A mutagenesis study of a catalytic antibody. United States: N. p., 1991. Web. doi:10.1073/pnas.88.1.58.
Jackson, D.Y., Prudent, J.R., Baldwin, E.P., & Schultz, P.G. A mutagenesis study of a catalytic antibody. United States. doi:10.1073/pnas.88.1.58.
Jackson, D.Y., Prudent, J.R., Baldwin, E.P., and Schultz, P.G. Tue . "A mutagenesis study of a catalytic antibody". United States. doi:10.1073/pnas.88.1.58.
@article{osti_5044075,
title = {A mutagenesis study of a catalytic antibody},
author = {Jackson, D.Y. and Prudent, J.R. and Baldwin, E.P. and Schultz, P.G.},
abstractNote = {The authors have generated seven site-specific mutations in the genes encoding the variable region of the heavy chain domain (V{sub H}) of the phosphocholine-binding antibody S107.S107 is a member of a family of well-characterized highly homologous antibodies that bind phosphorylcholine mono- and diesters. Two of these antibodies, MOPC-167 and T15, have previously been shown to catalyze the hydrolysis of 4-nitrophenyl N-trimethylammonioethyl carbonate. Two conserved heavy-chain residues, Tyr-33 and Arg-52, were postulated to be involved in binding and hydrolysis of 4-nitrophenylcholine carbonate esters. To more precisely define the catalytic roles of these residues, three Arg-52 mutants (R52K, R52Q, R52C) and four Tyr-33 mutants (Y33H, Y33F, Y33E, Y33D) of antibody S107 were generated. The genes encoding the V{sub H} binding domain of S107 were inserted into plasmid pUC-fl, and in vitro mutagenesis was performed. These results not only demonstrate the importance of electrostatic interactions in catalysis by antibody S107 but also show that catalytic side chains can be introduced into antibodies to enhance their catalytic efficiency.},
doi = {10.1073/pnas.88.1.58},
journal = {Proceedings of the National Academy of Sciences of the United States of America; (United States)},
issn = {0027-8424},
number = ,
volume = 88:1,
place = {United States},
year = {1991},
month = {1}
}