Transition state stabilization and binding energy use by catalytic antibody 17E8: Studies with alternative substrates

Herschel Wade, Thomas (Tom) Scanlan

Research output: Contribution to journalArticle

Abstract

The esterolytic antibody 17128 catalyzes the hydrolysis of n-formyl norleucine phenyl ester. The crystal structure of the antibody Fab fragment complexed to the transition state analog has been solved to 2.5 ± (Zhou et al, Science(1994)265,1059). The structure contains both an amino acid sidechain binding pocket and a pocket for the phenyl leaving group. These pockets seem to be the main determinants for binding to the transition state attalog and, presumably, the transition state. Previous kinetic results from our laboratory indicate that the noncovalent interactions between the Sl-pocket and the Pl-side chain of the substrate are essential for catalysis (Wade & Scanlan. JACS(1996)118,6510). In-this study, protein-substrate/transition state inter actions (S1-P1) were probed by a series of alternative substrates, substrate analogs, and transition state analogs to investigate the chemical requirements for substrate and transition state recognition. Several specific issues addressed by the alternative substrates include: number of VDW contacts, hydrophobic nature of the contacts, and the conformational requirements of the side chain. The results from these studies reveal the nature of the S1-P1 interactions and suggest how the binding energy from interactions involving this "less-evolved" protein pocket/active site is manifested in catalysis. This work is supported by the National Institutes of Health and the National Science Foundation.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - 1997
Externally publishedYes

Fingerprint

Catalytic Antibodies
Binding energy
Catalysis
catalytic activity
Stabilization
Norleucine
National Institutes of Health
Immunoglobulin Fragments
Immunoglobulin Fab Fragments
antibodies
energy
National Institutes of Health (U.S.)
Substrates
crystal structure
active sites
Catalytic Domain
Esters
Proteins
Hydrolysis
proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

@article{e510ed7817ef4189a967f32b3e8e3ff8,
title = "Transition state stabilization and binding energy use by catalytic antibody 17E8: Studies with alternative substrates",
abstract = "The esterolytic antibody 17128 catalyzes the hydrolysis of n-formyl norleucine phenyl ester. The crystal structure of the antibody Fab fragment complexed to the transition state analog has been solved to 2.5 ± (Zhou et al, Science(1994)265,1059). The structure contains both an amino acid sidechain binding pocket and a pocket for the phenyl leaving group. These pockets seem to be the main determinants for binding to the transition state attalog and, presumably, the transition state. Previous kinetic results from our laboratory indicate that the noncovalent interactions between the Sl-pocket and the Pl-side chain of the substrate are essential for catalysis (Wade & Scanlan. JACS(1996)118,6510). In-this study, protein-substrate/transition state inter actions (S1-P1) were probed by a series of alternative substrates, substrate analogs, and transition state analogs to investigate the chemical requirements for substrate and transition state recognition. Several specific issues addressed by the alternative substrates include: number of VDW contacts, hydrophobic nature of the contacts, and the conformational requirements of the side chain. The results from these studies reveal the nature of the S1-P1 interactions and suggest how the binding energy from interactions involving this {"}less-evolved{"} protein pocket/active site is manifested in catalysis. This work is supported by the National Institutes of Health and the National Science Foundation.",
author = "Herschel Wade and Scanlan, {Thomas (Tom)}",
year = "1997",
language = "English (US)",
volume = "11",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "9",

}

TY - JOUR

T1 - Transition state stabilization and binding energy use by catalytic antibody 17E8

T2 - Studies with alternative substrates

AU - Wade, Herschel

AU - Scanlan, Thomas (Tom)

PY - 1997

Y1 - 1997

N2 - The esterolytic antibody 17128 catalyzes the hydrolysis of n-formyl norleucine phenyl ester. The crystal structure of the antibody Fab fragment complexed to the transition state analog has been solved to 2.5 ± (Zhou et al, Science(1994)265,1059). The structure contains both an amino acid sidechain binding pocket and a pocket for the phenyl leaving group. These pockets seem to be the main determinants for binding to the transition state attalog and, presumably, the transition state. Previous kinetic results from our laboratory indicate that the noncovalent interactions between the Sl-pocket and the Pl-side chain of the substrate are essential for catalysis (Wade & Scanlan. JACS(1996)118,6510). In-this study, protein-substrate/transition state inter actions (S1-P1) were probed by a series of alternative substrates, substrate analogs, and transition state analogs to investigate the chemical requirements for substrate and transition state recognition. Several specific issues addressed by the alternative substrates include: number of VDW contacts, hydrophobic nature of the contacts, and the conformational requirements of the side chain. The results from these studies reveal the nature of the S1-P1 interactions and suggest how the binding energy from interactions involving this "less-evolved" protein pocket/active site is manifested in catalysis. This work is supported by the National Institutes of Health and the National Science Foundation.

AB - The esterolytic antibody 17128 catalyzes the hydrolysis of n-formyl norleucine phenyl ester. The crystal structure of the antibody Fab fragment complexed to the transition state analog has been solved to 2.5 ± (Zhou et al, Science(1994)265,1059). The structure contains both an amino acid sidechain binding pocket and a pocket for the phenyl leaving group. These pockets seem to be the main determinants for binding to the transition state attalog and, presumably, the transition state. Previous kinetic results from our laboratory indicate that the noncovalent interactions between the Sl-pocket and the Pl-side chain of the substrate are essential for catalysis (Wade & Scanlan. JACS(1996)118,6510). In-this study, protein-substrate/transition state inter actions (S1-P1) were probed by a series of alternative substrates, substrate analogs, and transition state analogs to investigate the chemical requirements for substrate and transition state recognition. Several specific issues addressed by the alternative substrates include: number of VDW contacts, hydrophobic nature of the contacts, and the conformational requirements of the side chain. The results from these studies reveal the nature of the S1-P1 interactions and suggest how the binding energy from interactions involving this "less-evolved" protein pocket/active site is manifested in catalysis. This work is supported by the National Institutes of Health and the National Science Foundation.

UR - http://www.scopus.com/inward/record.url?scp=33750125033&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750125033&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:33750125033

VL - 11

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 9

ER -