Substituent Effects on an Antibody-Catalyzed Hydrolysis of Phenyl Esters: Further Evidence for an Acyl-Antibody Intermediate

Richard A. Gibbs, Patricia A. Benkovic, Kim D. Janda, Richard A. Lerner, Stephen J. Benkovic

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

The hydrolysis of a series of para-substituted phenyl esters (2 and 6a-d) by the monoclonal catalytic antibody NPN43C9 has been investigated. The apparent pKa observed in the kcat/Km-pH profiles shifts from 8.9 (p-nitrophenyl ester) up to a maximum of 9.5 (p-methylphenyl ester). A correlation of the antibody-catalyzed rates of hydrolysis of the esters versus the σ parameter affords a ρ value of 2.3, indicative of a hydrolytic mechanism proceeding via the attack of a neutral nitrogen nucleophile and contrasting with the low ρ value expected for general base catalyzed hydrolysis. There was an inverse solvent deuterium isotope effect on the antibody-catalyzed hydrolysis of the p-chlorophenyl ester at pH > 10. These observations are consistent with our previous proposal (Benkovic, et al. Science 1990, 250, 1135) that this antibody employs a multistep kinetic pathway which involves the formation of a covalent acyl-antibody intermediate.

Original languageEnglish (US)
Pages (from-to)3528-3534
Number of pages7
JournalJournal of the American Chemical Society
Volume114
Issue number9
DOIs
StatePublished - Apr 1 1992

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Antibodies
Hydrolysis
Esters
Catalytic Antibodies
Nucleophiles
Deuterium
Isotopes
Nitrogen
Monoclonal Antibodies
Kinetics

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Gibbs, Richard A. ; Benkovic, Patricia A. ; Janda, Kim D. ; Lerner, Richard A. ; Benkovic, Stephen J. / Substituent Effects on an Antibody-Catalyzed Hydrolysis of Phenyl Esters : Further Evidence for an Acyl-Antibody Intermediate. In: Journal of the American Chemical Society. 1992 ; Vol. 114, No. 9. pp. 3528-3534.
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Substituent Effects on an Antibody-Catalyzed Hydrolysis of Phenyl Esters : Further Evidence for an Acyl-Antibody Intermediate. / Gibbs, Richard A.; Benkovic, Patricia A.; Janda, Kim D.; Lerner, Richard A.; Benkovic, Stephen J.

In: Journal of the American Chemical Society, Vol. 114, No. 9, 01.04.1992, p. 3528-3534.

Research output: Contribution to journalArticle

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