Probing the structure of HIV-1 Rev by protein footprinting of multiple monoclonal antibody-binding sites

FEBS Lett. 1997 Sep 1;414(1):50-4. doi: 10.1016/s0014-5793(97)00988-5.

Abstract

Human immunodeficiency virus type 1 (HIV-1) Rev is a small RNA-binding protein which is essential for viral replication. To investigate the structure of Rev we have mapped the binding sites of a panel of monoclonal antibodies (mAb) by protein footprinting and identified a mAb protecting amino acids within both the N- and C-terminal parts of Rev. Our mapping results support a previously proposed structure (Auer et al., Biochemistry, 33 (1994) 2988-2996) predicting that a helix-loop-helix motif in Rev brings the termini of the protein into proximity. Furthermore, we demonstrate that the binding sites mapped by protein footprinting are in agreement with conventional epitope mapping results and that this technique provides an advantageous strategy for mapping discontinuous sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / metabolism
  • Binding Sites
  • Bromelains / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Endopeptidase K / metabolism
  • Epitope Mapping
  • Gene Products, rev / chemistry*
  • Gene Products, rev / immunology
  • Gene Products, rev / metabolism
  • HIV-1 / chemistry*
  • Helix-Loop-Helix Motifs
  • Humans
  • Models, Molecular
  • Protein Conformation
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / immunology
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • rev Gene Products, Human Immunodeficiency Virus

Substances

  • Antibodies, Monoclonal
  • Gene Products, rev
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins
  • rev Gene Products, Human Immunodeficiency Virus
  • Bromelains
  • Endopeptidase K