Probing the structure of HIV-1 Rev by protein footprinting of multiple monoclonal antibody-binding sites.
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.
Jensen
T H
TH
Department of Molecular and Structural Biology, University of Aarhus, Aarhus C, Denmark.
Jensen
A
A
Szilvay
A M
AM
Kjems
J
J
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D001665:Binding Sites; D001963:Bromelains / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000378:metabolism; D018604:Epitope Mapping; D015694:Gene Products, rev / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D015497:HIV-1 / Q000737:chemistry*; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D054321:rev Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D015694:Gene Products, rev; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D001963:Bromelains; D019286:Endopeptidase K