Characterization of the vasoactive intestinal peptide (VIP) binding sites: a biochemical and an immunological approach.
The initial event of VIP action is its interaction with a specific receptor at the surface of a target cell. The understanding of the fine mechanism of action of VIP requires the characterization and of course the purification of the receptor. The understanding of the mechanism which regulates the number of receptor sites at the cell surface, if it occurs, is also a way to characterize the properties of VIP receptor. In this paper we report a number of data which represent several attempts to characterize VIP receptor in a human colonic adenocarcinoma cell (HT 29 cells). We have characterized a monoclonal antibody which partially inhibits 125I-VIP binding to HT 29 cells. We have specifically cross-linked, on intact HT 29 cells, a major polypeptide of Mr-64,000 with 125I-VIP using DTSP or DSS as cross-linking reagents. This polypeptide behaves like a high affinity binding site for VIP. We have demonstrated that VIP is rapidly internalized in HT 29 cells (in less than 10 minutes) and that simultaneously VIP receptors were no more detectable on the cell surface by cross-linking experiments. This suggests that VIP is internalized together with its receptor.
D000230:Adenocarcinoma / Q000378:metabolism; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002460:Cell Line; D003110:Colonic Neoplasms / Q000378:metabolism; D003432:Cross-Linking Reagents; D006801:Humans; D007120:Immunochemistry; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018005:Receptors, Vasoactive Intestinal Peptide; D014660:Vasoactive Intestinal Peptide / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D010455:Peptides; D011956:Receptors, Cell Surface; D018005:Receptors, Vasoactive Intestinal Peptide; D014660:Vasoactive Intestinal Peptide