Identification of binding proteins for pertussis toxin on pancreatic β cell-derived insulin-secreting cells

The ability of pertussis toxin (PT) to recognize and bind to surface proteins on cells derived from pancreatic insulin-secreting β cells and α cell-like glucagon-producing cells was investigated employing HIT-T15 ( β cell-derived) and In-R1-G9 (α cell-like) cell lines. PT recognition of membrane bin...

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Bibliographic Details
Published in:	Microbial pathogenesis Vol. 18; no. 3; pp. 173 - 185
Main Authors:	Bay⫢, Ali el, Linnemann, Ruth, von Olleschik-Elbheim, Lars, Alexander Schmidt, M.
Format:	Journal Article
Language:	English
Published:	Oxford Elsevier India Pvt Ltd 01-03-1995 Elsevier
Subjects:	Animals Bacteriology Binding proteins Biological and medical sciences Bordetella pertussis Carrier Proteins - chemistry Carrier Proteins - metabolism Cell Line CHO Cells Cricetinae Fundamental and applied biological sciences. Psychology Insulin - metabolism Insulin Secretion Insulin-secreting β-cells Islets of Langerhans - metabolism Lectin-binding Lectins - metabolism Microbiology Molecular Weight Neuraminidase Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains Pertussis Toxin Plant Lectins Receptors, Cell Surface - metabolism Ribosome Inactivating Proteins Sialic Acids - metabolism Virulence Factors, Bordetella - metabolism Lectin-binding Binding proteins Pertussis toxin Insulin-secreting β-cells Langerhans islet Rodentia Host agent relation Islet activating protein Glycoproteins Bordetella pertussis Binding protein Toxin Vertebrata Cell line Mammalia B-Cell Bacteria Pancreas Hamster Biological receptor
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Summary:	The ability of pertussis toxin (PT) to recognize and bind to surface proteins on cells derived from pancreatic insulin-secreting β cells and α cell-like glucagon-producing cells was investigated employing HIT-T15 ( β cell-derived) and In-R1-G9 (α cell-like) cell lines. PT recognition of membrane binding proteins on HIT-T15 and In-R1-G9 cells was first assessed with immunoflourescence microscopy in tissue culture. Both cell lines were equally well recognized by PT. N-octylglucoside extracts of whole cells and isolated membranes were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and blotted onto nitrocellulose membranes. PT, the B—oligomer, or the isolated PT dimers S2–S4 and S3–S4 recognized distinct proteins in HIT-T15 and In-R1-G9 cells of about 220 kDa. Recognition by the sialic acid specific Sambucus nigrica lectin identified these proteins as sialoglycoproteins. Incubation of the blotted membrane proteins with sialidase or pretreatment of PT with anti-PT polyclonal antibodies abolished the recognition and binding of these proteins by PT. To demonstrate that these glycoproteins are also able to transduce PT mediated effects and thus might serve as PT binding proteins, the stimulation of insulin secretion in HIT-T15 cells was assessed. As the secretion of insulin in HIT-T15 cells increased about 30% upon interaction with PT it was concluded that these glycoproteins are indeed functional as PT receptors.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0882-4010 1096-1208
DOI:	10.1016/S0882-4010(95)90031-4