Catalytic Antibodies with Acyl-Transfer Capabilities

Mechanistic and Kinetic Investigations

Kim D. Janda, Jon A. Ashley, Teresa M. Jones, Donald A. McLeod, Diane M. Schloeder, Michael I. Weinhouse, Richard A. Lerner, Richard A. Gibbs, Patricia Ann Benkovic, Riet Hilhorst, Stephen Benkovic

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Antibodies have been shown to catalyze acyl-transfer reactions. The processes by which they perform such tasks have often been postulated but largely remain unknown. An extended study is presented on three different monoclonal antibodies that catalyze the hydrolysis of an alkyl ester and an aryl amide bond. Antibodies 2H6 and 21H3 catalyze the hydrolysis of an unactivated benzyl ester and show exquisite specificity for substrates with either the R or S configuration, respectively, while 43C9 catalyzes the cleavage of a p-nitroanilide amide bond. New substrates were synthesized and buffer-assisted reactions were employed to determine antibody-substrate fidelity. Oxygen-18 incorporation experiments were performed providing evidence that these antibody-mediated reactions proceed through attack at the acyl carbonyl, and excluding the possibility of an SN2 displacement mechanism for the ester hydrolysis reaction. A pH-rate profile study in protium and deuterium oxide was performed on antibody 43C9. This revealed an apparent pKa of ~9 involved in catalysis, but both the presence and absence of a solvent isotope effect in the pH-dependent and -independent regions suggested a multistep reaction pathway may be operative.

Original languageEnglish (US)
Pages (from-to)291-297
Number of pages7
JournalJournal of the American Chemical Society
Volume113
Issue number1
DOIs
StatePublished - Jan 1 1991

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Catalytic Antibodies
Antibodies
Kinetics
Hydrolysis
Esters
Amides
Substrates
Deuterium Oxide
Heavy water
Monoclonal antibodies
Substrate Specificity
Catalysis
Isotopes
Buffers
Monoclonal Antibodies
Oxygen

All Science Journal Classification (ASJC) codes

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

Cite this

Janda, K. D., Ashley, J. A., Jones, T. M., McLeod, D. A., Schloeder, D. M., Weinhouse, M. I., ... Benkovic, S. (1991). Catalytic Antibodies with Acyl-Transfer Capabilities: Mechanistic and Kinetic Investigations. Journal of the American Chemical Society, 113(1), 291-297. https://doi.org/10.1021/ja00001a042
Janda, Kim D. ; Ashley, Jon A. ; Jones, Teresa M. ; McLeod, Donald A. ; Schloeder, Diane M. ; Weinhouse, Michael I. ; Lerner, Richard A. ; Gibbs, Richard A. ; Benkovic, Patricia Ann ; Hilhorst, Riet ; Benkovic, Stephen. / Catalytic Antibodies with Acyl-Transfer Capabilities : Mechanistic and Kinetic Investigations. In: Journal of the American Chemical Society. 1991 ; Vol. 113, No. 1. pp. 291-297.
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abstract = "Antibodies have been shown to catalyze acyl-transfer reactions. The processes by which they perform such tasks have often been postulated but largely remain unknown. An extended study is presented on three different monoclonal antibodies that catalyze the hydrolysis of an alkyl ester and an aryl amide bond. Antibodies 2H6 and 21H3 catalyze the hydrolysis of an unactivated benzyl ester and show exquisite specificity for substrates with either the R or S configuration, respectively, while 43C9 catalyzes the cleavage of a p-nitroanilide amide bond. New substrates were synthesized and buffer-assisted reactions were employed to determine antibody-substrate fidelity. Oxygen-18 incorporation experiments were performed providing evidence that these antibody-mediated reactions proceed through attack at the acyl carbonyl, and excluding the possibility of an SN2 displacement mechanism for the ester hydrolysis reaction. A pH-rate profile study in protium and deuterium oxide was performed on antibody 43C9. This revealed an apparent pKa of ~9 involved in catalysis, but both the presence and absence of a solvent isotope effect in the pH-dependent and -independent regions suggested a multistep reaction pathway may be operative.",
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Janda, KD, Ashley, JA, Jones, TM, McLeod, DA, Schloeder, DM, Weinhouse, MI, Lerner, RA, Gibbs, RA, Benkovic, PA, Hilhorst, R & Benkovic, S 1991, 'Catalytic Antibodies with Acyl-Transfer Capabilities: Mechanistic and Kinetic Investigations', Journal of the American Chemical Society, vol. 113, no. 1, pp. 291-297. https://doi.org/10.1021/ja00001a042

Catalytic Antibodies with Acyl-Transfer Capabilities : Mechanistic and Kinetic Investigations. / Janda, Kim D.; Ashley, Jon A.; Jones, Teresa M.; McLeod, Donald A.; Schloeder, Diane M.; Weinhouse, Michael I.; Lerner, Richard A.; Gibbs, Richard A.; Benkovic, Patricia Ann; Hilhorst, Riet; Benkovic, Stephen.

In: Journal of the American Chemical Society, Vol. 113, No. 1, 01.01.1991, p. 291-297.

Research output: Contribution to journalArticle

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T1 - Catalytic Antibodies with Acyl-Transfer Capabilities

T2 - Mechanistic and Kinetic Investigations

AU - Janda, Kim D.

AU - Ashley, Jon A.

AU - Jones, Teresa M.

AU - McLeod, Donald A.

AU - Schloeder, Diane M.

AU - Weinhouse, Michael I.

AU - Lerner, Richard A.

AU - Gibbs, Richard A.

AU - Benkovic, Patricia Ann

AU - Hilhorst, Riet

AU - Benkovic, Stephen

PY - 1991/1/1

Y1 - 1991/1/1

N2 - Antibodies have been shown to catalyze acyl-transfer reactions. The processes by which they perform such tasks have often been postulated but largely remain unknown. An extended study is presented on three different monoclonal antibodies that catalyze the hydrolysis of an alkyl ester and an aryl amide bond. Antibodies 2H6 and 21H3 catalyze the hydrolysis of an unactivated benzyl ester and show exquisite specificity for substrates with either the R or S configuration, respectively, while 43C9 catalyzes the cleavage of a p-nitroanilide amide bond. New substrates were synthesized and buffer-assisted reactions were employed to determine antibody-substrate fidelity. Oxygen-18 incorporation experiments were performed providing evidence that these antibody-mediated reactions proceed through attack at the acyl carbonyl, and excluding the possibility of an SN2 displacement mechanism for the ester hydrolysis reaction. A pH-rate profile study in protium and deuterium oxide was performed on antibody 43C9. This revealed an apparent pKa of ~9 involved in catalysis, but both the presence and absence of a solvent isotope effect in the pH-dependent and -independent regions suggested a multistep reaction pathway may be operative.

AB - Antibodies have been shown to catalyze acyl-transfer reactions. The processes by which they perform such tasks have often been postulated but largely remain unknown. An extended study is presented on three different monoclonal antibodies that catalyze the hydrolysis of an alkyl ester and an aryl amide bond. Antibodies 2H6 and 21H3 catalyze the hydrolysis of an unactivated benzyl ester and show exquisite specificity for substrates with either the R or S configuration, respectively, while 43C9 catalyzes the cleavage of a p-nitroanilide amide bond. New substrates were synthesized and buffer-assisted reactions were employed to determine antibody-substrate fidelity. Oxygen-18 incorporation experiments were performed providing evidence that these antibody-mediated reactions proceed through attack at the acyl carbonyl, and excluding the possibility of an SN2 displacement mechanism for the ester hydrolysis reaction. A pH-rate profile study in protium and deuterium oxide was performed on antibody 43C9. This revealed an apparent pKa of ~9 involved in catalysis, but both the presence and absence of a solvent isotope effect in the pH-dependent and -independent regions suggested a multistep reaction pathway may be operative.

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