Monoclonal Antibodies against HIV Type 1 Integrase: Clues to Molecular Structure
- 1 September 1994
- journal article
- research article
- Published by Mary Ann Liebert Inc in AIDS Research and Human Retroviruses
- Vol. 10 (9), 1105-1115
- https://doi.org/10.1089/aid.1994.10.1105
Abstract
Eleven murine hybridoma clones were selected for their ability to produce anti-HIV-1 integrase (IN) antibodies. Competition and epitope mapping studies allowed segregation of the monoclonal antibodies (MAbs) into four distinct classes. The five MAbs that comprise the first class showed high affinity for epitopes within an N-terminal domain of 58 amino acids that includes a conserved zinc finger motif. The second class, with two MAbs, showed high affinity for epitopes within 29 amino acids at the C terminus. Another two MAbs, which constitute the third class, displayed moderate affinities for epitopes that mapped to regions within the highly conserved catalytic core referred to as the D,D(35)E domain. One of these MAbs showed significant cross-reactivity with HIV-2 IN and weak, but detectable, cross-reactivity with RSV IN. The remaining two MAbs, which comprise the fourth class, exhibited fairly low binding affinities and appeared to recognize epitopes in the zinc finger motif domain as well as the C-terminal half of the IN protein. The MAbs can be used for immunoprecipitation and immunoblotting procedures as well as for purification of HIV-1 IN protein by affinity chromatography. We show that several can also be used to immunostain viral IN sequences in HIV-1-infected T cells, presumably as a component of Gag-Pol precursors. Finally, analysis of our mapping and competition data suggests a structure for mature IN in which the C terminus approaches the central core domain, and the N and C termini touch or are proximal to each other. These MAbs should prove useful for further analyses of the structure and function of IN both in vitro and in vivo.Keywords
This publication has 30 references indexed in Scilit:
- Molecular mechanism of retroviral DNA integrationPharmacology & Therapeutics, 1994
- THE RETROVIRAL ENZYMESAnnual Review of Biochemistry, 1994
- Crystal structure of human immunodeficiency virus type 1 reverse transcriptase complexed with double-stranded DNA at 3.0 A resolution shows bent DNA.Proceedings of the National Academy of Sciences, 1993
- Mapping of Immunodominant Epitopes of the HIV-1 and HIV-2 Integrase Proteins by Recombinant Proteins and Synthetic PeptidesAIDS Research and Human Retroviruses, 1992
- Identification of amino acid residues critical for endonuclease and integration activities of HIV-1 IN protein in VitroVirology, 1992
- Localization of DNA Binding Activity of HIV-1 Integrase to the C-Terminal Half of the ProteinAIDS Research and Human Retroviruses, 1992
- Preparation and crystallization of a human immunodeficiency virus p24-Fab complex.Proceedings of the National Academy of Sciences, 1990
- Human immunodeficiency virus integration protein expressed in Escherichia coli possesses selective DNA cleaving activity.Proceedings of the National Academy of Sciences, 1990
- Requirement of the avian retrovirus pp32 DNA binding protein domain for replicationVirology, 1984
- A better cell line for making hybridomas secreting specific antibodiesNature, 1978