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1996-02-26
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Document 0467
DOCN M9620467
TI Conformational changes between human immunodeficiency virus type 1
nucleocapsid protein NCp7 and its precursor NCp15 as detected by
anti-NCp7 monoclonal antibodies.
DT 9602
AU Tanchou V; Delaunay T; Bodeus M; Roques B; Darlix JL; Benarous R;
Signalisation, Inflammation et Transformation Cellulaire, U332; INSERM,
Paris, France.
SO J Gen Virol. 1995 Oct;76 ( Pt 10):2457-66. Unique Identifier : AIDSLINE
MED/96030855
AB The nucleocapsid protein NCp15 of human immunodeficiency virus type 1
(HIV-1) is a small basic protein with two zinc fingers. It is required
for virion morphogenesis and synthesis of proviral DNA. As the first
step in our study of the structural domains involved in the various
functions of this protein, 18 monoclonal antibodies (MAbs) were
isolated. The epitopes of NCp7 recognized by the MAbs were mapped using
synthetic peptides representing overlapping sequences and truncated
forms of NCp7. These anti-NCp7 MAbs were investigated by ELISA and
real-time biospecific interaction analysis (BIAcore). Five classes of
anti-NCp7 MAbs were characterized. Three classes (14 MAbs) were directed
against continuous epitopes, one in the N-terminal part, another next to
the second zinc finger and the third in the C-terminal part of the
protein. Two other classes comprised four MAbs reacting only with the
entire NCp7 and not with any of the small overlapping peptides used,
suggesting that these MAbs were directed against conformational
epitopes. The anti-NCp7 MAbs directed against linear epitopes were able
to react efficiently with both NCp7 and NCp15, the NCp7 precursor,
whereas the anti-NCp7 MAbs directed against conformational epitopes did
not react with NCp15. Interestingly, most of the anti-NCp7 MAbs directed
against conformational epitopes were capable of inhibiting the tight
interaction between NCp7 and the HIV-1 replication primer tRNA(Lys,3).
In contrast, most of the MAbs directed against linear epitopes did not
inhibit this interaction.
DE Amino Acid Sequence Animal *Antibodies, Monoclonal Antigen-Antibody
Reactions Binding, Competitive Biosensors
Capsid/*CHEMISTRY/IMMUNOLOGY Epitope Mapping Female Gene Products,
gag/*CHEMISTRY/IMMUNOLOGY Human *HIV Antibodies HIV-1/*IMMUNOLOGY
Mice Mice, Inbred BALB C Molecular Sequence Data Peptides/CHEMICAL
SYNTHESIS *Protein Conformation Protein Precursors/CHEMISTRY Rats
RNA, Transfer, Lys/ANALYSIS Support, Non-U.S. Gov't Zinc
Fingers/IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).