Crystal structure of the complex of a catalytic antibody Fab fragment with a transition state analog: structural similarities in esterase-like catalytic antibodies

Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11721-5. doi: 10.1073/pnas.92.25.11721.

Abstract

The x-ray structure of the complex of a catalytic antibody Fab fragment with a phosphonate transition-state analog has been determined. The antibody (CNJ206) catalyzes the hydrolysis of p-nitrophenyl esters with significant rate enhancement and substrate specificity. Comparison of this structure with that of the uncomplexed Fab fragment suggests hapten-induced conformational changes: the shape of the combining site changes from a shallow groove in the uncomplexed Fab to a deep pocket where the hapten is buried. Three hydrogen-bond donors appear to stabilize the charged phosphonate group of the hapten: two NH groups of the heavy (H) chain complementarity-determining region 3 (H3 CDR) polypeptide chain and the side-chain of histidine-H35 in the H chain (His-H35) in the H1 CDR. The combining site shows striking structural similarities to that of antibody 17E8, which also has esterase activity. Both catalytic antibody ("abzyme") structures suggest that oxyanion stabilization plays a significant role in their rate acceleration. Additional catalytic groups that improve efficiency are not necessarily induced by the eliciting hapten; these groups may occur because of the variability in the combining sites of different monoclonal antibodies that bind to the same hapten.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Catalytic / chemistry*
  • Antibodies, Catalytic / metabolism
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / metabolism
  • Binding Sites
  • Computer Simulation
  • Crystallography
  • Esterases / chemistry*
  • Esterases / metabolism
  • Haptens / chemistry
  • Haptens / metabolism
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Fab Fragments / metabolism
  • Kinetics
  • Models, Molecular
  • Nitrophenols / chemistry
  • Nitrophenols / immunology
  • Protein Conformation
  • Substrate Specificity
  • Synchrotrons

Substances

  • Antibodies, Catalytic
  • Antibodies, Monoclonal
  • Haptens
  • Immunoglobulin Fab Fragments
  • Nitrophenols
  • Esterases
  • 4-nitrophenol