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

FEBS Letters

Kjems, Jørgen²  Jensen, Torben Heick²  Szilvay, Anne Marie¹  Jensen, Allan² 

[1]Centre for Research in Virology, The Gade Institute, Department of Microbiology and Immunology, University of Bergen, Bergen, Norway

[2]Department of Molecular and Structural Biology, University of Aarhus, C.F. Møllers Allé Building 130, DK-8000 Aarhus C, Denmark

关键词: HIV;    Rev;    Protein footprinting;    Epitope mapping;    Conformational epitope;    HIV-1;    human immunodeficiency virus type 1;    aa;    amino acid(s);    NMR;    nuclear magnetic resonance;    mAb;    monoclonal antibody;    HMK;    heart muscle kinase;    GST;    glutathione S-transferase;    NES;    nuclear export signal;   

DOI  :  10.1016/S0014-5793(97)00988-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown