A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla)
ABSTRACT Using labelled ligand-binding methods, previous studies have identified specific angiotensin II receptors (Ang II-Rs) in eel liver, kidney and intestine membranes. Isoelectric focusing on polyacrylamide gels also showed that there are two Ang II-R isoforms in eel liver, focusing at isoelect...
Published in: | Journal of Molecular Endocrinology |
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Bioscientifica
1996
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crbioscientif:10.1677/jme.0.0160045 2023-11-12T04:01:15+01:00 A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) Marsigliante, S Muscella, A Vilella, S Nicolardi, G Ingrosso, L Ciardo, V Zonno, V Vinson, G P Ho, M M Storelli, C 1996 http://dx.doi.org/10.1677/jme.0.0160045 https://jme.bioscientifica.com/view/journals/jme/16/1/jme_16_1_006.xml https://jme.bioscientifica.com/downloadpdf/journals/jme/16/1/jme_16_1_006.xml unknown Bioscientifica Journal of Molecular Endocrinology volume 16, issue 1, page 45-56 ISSN 0952-5041 1479-6813 Endocrinology Molecular Biology journal-article 1996 crbioscientif https://doi.org/10.1677/jme.0.0160045 2023-10-20T08:24:14Z ABSTRACT Using labelled ligand-binding methods, previous studies have identified specific angiotensin II receptors (Ang II-Rs) in eel liver, kidney and intestine membranes. Isoelectric focusing on polyacrylamide gels also showed that there are two Ang II-R isoforms in eel liver, focusing at isoelectric points (pI) 6·5 and 6·7. These may have different functions. In contrast, eel enterocyte plasma membrane and renal brush border membranes contain only the pI 6·5 form. To characterize the eel receptors more fully, a newly developed monoclonal antibody (6313/G2) which selectively recognizes the AT 1 subtype of mammalian Ang II-R was used. In ligand-binding experiments, the preincubation of eel liver membranes with 6313/G2 antibody eliminated the specific [3,5- 3 H]Tyr 4 -Ile 5 -Ang II binding. Moreover, Ang II—receptor complexes from solubilized liver membranes, which were immunoprecipitated by 6313/G2-coated beads, had a pI of 6·5. In immunoblotting experiments, the antibody recognized the isoform focusing at pI 6·5 in eel intestine and liver preparations, but not the liver pI 6·7 isoform. Immunoblotting of SDS gels showed that the antibody bound to a single protein of molecular mass of 75 kDa in eel liver, gill and kidney and to a doublet of molecular mass of about 74 and 75 kDa in intestinal membrane preparations. Immunocytochemistry of paraffin-embedded and cryostat sections of eel liver, kidney, intestine and gill showed that antibody 6313/G2 bound to uniformly distributed intracellular sites and cell surface membranes in proximal tubular cells, absorptive intestinal cells, hepatocytes and chloride cells. It also stained endothelium and both the longitudinal and circular layers of smooth muscle cells in the intestine. The data suggest that the previously described Ang II-R from eel liver, kidney and intestine may be similar to the mammalian AT 1 subtype. Article in Journal/Newspaper Anguilla anguilla Bioscientifica (via Crossref) Journal of Molecular Endocrinology 16 1 45 56 |
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Bioscientifica (via Crossref) |
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topic |
Endocrinology Molecular Biology |
spellingShingle |
Endocrinology Molecular Biology Marsigliante, S Muscella, A Vilella, S Nicolardi, G Ingrosso, L Ciardo, V Zonno, V Vinson, G P Ho, M M Storelli, C A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) |
topic_facet |
Endocrinology Molecular Biology |
description |
ABSTRACT Using labelled ligand-binding methods, previous studies have identified specific angiotensin II receptors (Ang II-Rs) in eel liver, kidney and intestine membranes. Isoelectric focusing on polyacrylamide gels also showed that there are two Ang II-R isoforms in eel liver, focusing at isoelectric points (pI) 6·5 and 6·7. These may have different functions. In contrast, eel enterocyte plasma membrane and renal brush border membranes contain only the pI 6·5 form. To characterize the eel receptors more fully, a newly developed monoclonal antibody (6313/G2) which selectively recognizes the AT 1 subtype of mammalian Ang II-R was used. In ligand-binding experiments, the preincubation of eel liver membranes with 6313/G2 antibody eliminated the specific [3,5- 3 H]Tyr 4 -Ile 5 -Ang II binding. Moreover, Ang II—receptor complexes from solubilized liver membranes, which were immunoprecipitated by 6313/G2-coated beads, had a pI of 6·5. In immunoblotting experiments, the antibody recognized the isoform focusing at pI 6·5 in eel intestine and liver preparations, but not the liver pI 6·7 isoform. Immunoblotting of SDS gels showed that the antibody bound to a single protein of molecular mass of 75 kDa in eel liver, gill and kidney and to a doublet of molecular mass of about 74 and 75 kDa in intestinal membrane preparations. Immunocytochemistry of paraffin-embedded and cryostat sections of eel liver, kidney, intestine and gill showed that antibody 6313/G2 bound to uniformly distributed intracellular sites and cell surface membranes in proximal tubular cells, absorptive intestinal cells, hepatocytes and chloride cells. It also stained endothelium and both the longitudinal and circular layers of smooth muscle cells in the intestine. The data suggest that the previously described Ang II-R from eel liver, kidney and intestine may be similar to the mammalian AT 1 subtype. |
format |
Article in Journal/Newspaper |
author |
Marsigliante, S Muscella, A Vilella, S Nicolardi, G Ingrosso, L Ciardo, V Zonno, V Vinson, G P Ho, M M Storelli, C |
author_facet |
Marsigliante, S Muscella, A Vilella, S Nicolardi, G Ingrosso, L Ciardo, V Zonno, V Vinson, G P Ho, M M Storelli, C |
author_sort |
Marsigliante, S |
title |
A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) |
title_short |
A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) |
title_full |
A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) |
title_fullStr |
A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) |
title_full_unstemmed |
A monoclonal antibody to mammalian angiotensin II AT1 receptor recognizes one of the angiotensin II receptor isoforms expressed by the eel (Anguilla anguilla) |
title_sort |
monoclonal antibody to mammalian angiotensin ii at1 receptor recognizes one of the angiotensin ii receptor isoforms expressed by the eel (anguilla anguilla) |
publisher |
Bioscientifica |
publishDate |
1996 |
url |
http://dx.doi.org/10.1677/jme.0.0160045 https://jme.bioscientifica.com/view/journals/jme/16/1/jme_16_1_006.xml https://jme.bioscientifica.com/downloadpdf/journals/jme/16/1/jme_16_1_006.xml |
genre |
Anguilla anguilla |
genre_facet |
Anguilla anguilla |
op_source |
Journal of Molecular Endocrinology volume 16, issue 1, page 45-56 ISSN 0952-5041 1479-6813 |
op_doi |
https://doi.org/10.1677/jme.0.0160045 |
container_title |
Journal of Molecular Endocrinology |
container_volume |
16 |
container_issue |
1 |
container_start_page |
45 |
op_container_end_page |
56 |
_version_ |
1782331949637959680 |