Generation of monoclonal antibodies specifically directed against the proximal zinc finger of HIV type 1 NCp7

Autor: Chang-Zhi Dong, Thierry Delaunay, Anne Caneparo, Bernard P. Roques, Hugues de Rocquigny
Přispěvatelé: ProdInra, Migration, Pharmacochimie moléculaire et structurale, Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), Génomique, développement et pouvoir pathogène (GD2P), Université Bordeaux Segalen - Bordeaux 2-Institut National de la Recherche Agronomique (INRA)
Rok vydání: 2000
Předmět:
[SDV]Life Sciences [q-bio]
Peptide
IMMUNOLOGIE
01 natural sciences
gag Gene Products
Human Immunodeficiency Virus
Epitope
Mice
ComputingMilieux_MISCELLANEOUS
Zinc finger
chemistry.chemical_classification
0303 health sciences
Mice
Inbred BALB C
biology
Antibodies
Monoclonal
3. Good health
Cell biology
[SDV] Life Sciences [q-bio]
Infectious Diseases
Female
Antibody
medicine.drug_class
Immunoprecipitation
Immunology
Molecular Sequence Data
Gene Products
gag
Enzyme-Linked Immunosorbent Assay
010402 general chemistry
Monoclonal antibody
Peptides
Cyclic
03 medical and health sciences
Viral Proteins
Capsid
Virology
medicine
Animals
Humans
Amino Acid Sequence
Protein Precursors
030304 developmental biology
Gene Products
vpr
HIV
Surface Plasmon Resonance
Molecular biology
Zinc finger nuclease
Precipitin Tests
0104 chemical sciences
Kinetics
chemistry
biology.protein
Capsid Proteins
Function (biology)
Epitope Mapping
Zdroj: AIDS Research and Human Retroviruses
AIDS Research and Human Retroviruses, Mary Ann Liebert, 2000, 16 (13), pp.1259-1267
ISSN: 0889-2229
2726-2731
Popis: The HIV-1 NCp7 contains two spatially close zinc fingers, required for the production of infectious particles. To investigate in more detail the function of the zinc finger domain, monoclonal antibodies were generated with a cyclic analog of the NCp7 proximal zinc finger. This analog was shown to bind zinc ions and to preserve the highly folded structure of the native peptide (Dong C-Z et al.: J Am Chem Soc 1995;117:2726-2731). We report here two monoclonal antibodies (2B10 and 4D3), which are the first monoclonal antibodies directed against CCHC NCp7 zinc fingers. Dot-blot experiments revealed that a few nanograms of synthetic NCp7 can be detected on a nitrocellulose membrane. Whereas 2B10 appears specific for an epitope located in sequence 19-27 of NCp7, 4D3 appears to be structurally specific. Immunocomplex affinities were evaluated, using BIAcore technology, to be up to 1 and 10 nM, respectively, for 2B10 and 4D3 in 100 mM NaCl. These antibodies were able to recognize NCp7 in the Gag polyprotein precursor and were shown to immunoprecipitate NCp7 from a cell supernatant. Moreover, NCp7-Vpr interaction mediated by the zinc fingers is inhibited by 2B10, emphasizing the role of these domains in the protein-protein complex. These results indicate that 2B10 and 4D3 behave as useful tools for studying both NC protein functions during the course of virion morphogenesis and the role played by its zinc finger domain at various steps in the retroviral life cycle.
Databáze: OpenAIRE
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