Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4
Feline immunodeficiency virus (FIV) and the T cell-tropic strains of human immunodeficiency virus type 1 (HIV-1) share the use of the chemokine receptor CXCR4 for cell entry. To study this process further we developed a cell surface binding assay based on the expression of a soluble version of the F...
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2014
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_08892229_v30_n3_p250_Gonzalez http://hdl.handle.net/20.500.12110/paper_08892229_v30_n3_p250_Gonzalez |
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paper:paper_08892229_v30_n3_p250_Gonzalez2023-06-08T15:47:05Z Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 González, Silvia Adriana Affranchino, José Luis binding protein chemokine receptor CXCR4 chimeric protein glycoprotein E1 Influenza virus hemagglutinin membrane protein monoclonal antibody plerixafor virus envelope protein article binding assay cell fusion cell surface controlled study electrophoretic mobility embryo enzyme linked immunosorbent assay Feline immunodeficiency virus flow cytometry human human cell Human immunodeficiency virus 1 immunoblotting polyacrylamide gel electrophoresis priority journal protein expression protein subunit quantitative analysis quantitative assay site directed mutagenesis surface property T lymphocyte target cell virus entry Western blotting Animals Cell Line env Gene Products, Human Immunodeficiency Virus Glycoproteins HIV-1 Humans Immunodeficiency Virus, Feline Receptors, CXCR4 Receptors, HIV Recombination, Genetic Viral Envelope Proteins Virus Attachment Feline immunodeficiency virus (FIV) and the T cell-tropic strains of human immunodeficiency virus type 1 (HIV-1) share the use of the chemokine receptor CXCR4 for cell entry. To study this process further we developed a cell surface binding assay based on the expression of a soluble version of the FIV SU C-terminally tagged with the influenza virus hemagglutinin epitope (HA). The specificity of the assay was demonstrated by the following evidence: (1) the SU-HA protein bound to HeLa cells that express CXCR4 but not to MDCK cells that lack this chemokine receptor; and (2) binding of the SU-HA to HeLa cells was blocked by incubation with the CXCR4 antagonist AMD3100 as well as with the anti-CXCR4 monoclonal antibody (MAb) 12G5. Deletion of the V3 region from the FIV SU glycoprotein abolished its ability to bind CXCR4-expressing cells. Remarkably, substitution of the V3 domain of the FIV SU by the equivalent region of the HIV-1 NL4-3 isolate resulted in efficient cell surface binding of the chimeric SU protein to CXCR4. Moreover, transfection of MDCK cells with a plasmid encoding human CXCR4 allowed the association of the chimeric SU-HA glycoprotein to the transfected cells. Interestingly, while cell binding of the chimeric FIV-HIV SU was inhibited by an anti-HIV-1 V3 MAb, its association with CXCR4 was found to be resistant to AMD3100. Of note, the chimeric FIV-HIV Env glycoprotein was capable of promoting CXCR4-dependent cell-to-cell fusion. © Copyright 2014, Mary Ann Liebert, Inc. 2014. Fil:González, S.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Affranchino, J.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2014 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_08892229_v30_n3_p250_Gonzalez http://hdl.handle.net/20.500.12110/paper_08892229_v30_n3_p250_Gonzalez |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
binding protein chemokine receptor CXCR4 chimeric protein glycoprotein E1 Influenza virus hemagglutinin membrane protein monoclonal antibody plerixafor virus envelope protein article binding assay cell fusion cell surface controlled study electrophoretic mobility embryo enzyme linked immunosorbent assay Feline immunodeficiency virus flow cytometry human human cell Human immunodeficiency virus 1 immunoblotting polyacrylamide gel electrophoresis priority journal protein expression protein subunit quantitative analysis quantitative assay site directed mutagenesis surface property T lymphocyte target cell virus entry Western blotting Animals Cell Line env Gene Products, Human Immunodeficiency Virus Glycoproteins HIV-1 Humans Immunodeficiency Virus, Feline Receptors, CXCR4 Receptors, HIV Recombination, Genetic Viral Envelope Proteins Virus Attachment |
| spellingShingle |
binding protein chemokine receptor CXCR4 chimeric protein glycoprotein E1 Influenza virus hemagglutinin membrane protein monoclonal antibody plerixafor virus envelope protein article binding assay cell fusion cell surface controlled study electrophoretic mobility embryo enzyme linked immunosorbent assay Feline immunodeficiency virus flow cytometry human human cell Human immunodeficiency virus 1 immunoblotting polyacrylamide gel electrophoresis priority journal protein expression protein subunit quantitative analysis quantitative assay site directed mutagenesis surface property T lymphocyte target cell virus entry Western blotting Animals Cell Line env Gene Products, Human Immunodeficiency Virus Glycoproteins HIV-1 Humans Immunodeficiency Virus, Feline Receptors, CXCR4 Receptors, HIV Recombination, Genetic Viral Envelope Proteins Virus Attachment González, Silvia Adriana Affranchino, José Luis Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 |
| topic_facet |
binding protein chemokine receptor CXCR4 chimeric protein glycoprotein E1 Influenza virus hemagglutinin membrane protein monoclonal antibody plerixafor virus envelope protein article binding assay cell fusion cell surface controlled study electrophoretic mobility embryo enzyme linked immunosorbent assay Feline immunodeficiency virus flow cytometry human human cell Human immunodeficiency virus 1 immunoblotting polyacrylamide gel electrophoresis priority journal protein expression protein subunit quantitative analysis quantitative assay site directed mutagenesis surface property T lymphocyte target cell virus entry Western blotting Animals Cell Line env Gene Products, Human Immunodeficiency Virus Glycoproteins HIV-1 Humans Immunodeficiency Virus, Feline Receptors, CXCR4 Receptors, HIV Recombination, Genetic Viral Envelope Proteins Virus Attachment |
| description |
Feline immunodeficiency virus (FIV) and the T cell-tropic strains of human immunodeficiency virus type 1 (HIV-1) share the use of the chemokine receptor CXCR4 for cell entry. To study this process further we developed a cell surface binding assay based on the expression of a soluble version of the FIV SU C-terminally tagged with the influenza virus hemagglutinin epitope (HA). The specificity of the assay was demonstrated by the following evidence: (1) the SU-HA protein bound to HeLa cells that express CXCR4 but not to MDCK cells that lack this chemokine receptor; and (2) binding of the SU-HA to HeLa cells was blocked by incubation with the CXCR4 antagonist AMD3100 as well as with the anti-CXCR4 monoclonal antibody (MAb) 12G5. Deletion of the V3 region from the FIV SU glycoprotein abolished its ability to bind CXCR4-expressing cells. Remarkably, substitution of the V3 domain of the FIV SU by the equivalent region of the HIV-1 NL4-3 isolate resulted in efficient cell surface binding of the chimeric SU protein to CXCR4. Moreover, transfection of MDCK cells with a plasmid encoding human CXCR4 allowed the association of the chimeric SU-HA glycoprotein to the transfected cells. Interestingly, while cell binding of the chimeric FIV-HIV SU was inhibited by an anti-HIV-1 V3 MAb, its association with CXCR4 was found to be resistant to AMD3100. Of note, the chimeric FIV-HIV Env glycoprotein was capable of promoting CXCR4-dependent cell-to-cell fusion. © Copyright 2014, Mary Ann Liebert, Inc. 2014. |
| author |
González, Silvia Adriana Affranchino, José Luis |
| author_facet |
González, Silvia Adriana Affranchino, José Luis |
| author_sort |
González, Silvia Adriana |
| title |
Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 |
| title_short |
Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 |
| title_full |
Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 |
| title_fullStr |
Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 |
| title_full_unstemmed |
Replacement of the V3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a T cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to CXCR4 |
| title_sort |
replacement of the v3 domain in the surface subunit of the feline immunodeficiency virus envelope glycoprotein with the equivalent region of a t cell-tropic human immunodeficiency virus type 1 results in a chimeric surface protein that efficiently binds to cxcr4 |
| publishDate |
2014 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_08892229_v30_n3_p250_Gonzalez http://hdl.handle.net/20.500.12110/paper_08892229_v30_n3_p250_Gonzalez |
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