Biological action and fate of photoaffinity-labelled insulin-receptor complexes.
Covalent linking of two photoactivatable insulin derivatives, B2-(2-nitro,4-azidophenylacetyl)-des-PheB1-insulin and B29-(2-nitro,4-azidophenylacetyl)-insulin to viable rat adipocytes gives a system, which contains a fixed stoichiometry between hormone and receptor. The biological signal of prolonged lipogenesis has been used to study several aspects of insulin binding and action: the role of the site of the crosslink between insulin and receptor, recognition of bound photoinsulin by anti-insulin antibodies, the half-life of the biologically active complex, the pH-dependence of the biological signal, and the possible role of internalization. Furthermore, the effect of trypsin on the insulin receptor, as well as the insulin-receptor complex, has been investigated and a refined model of the receptor is presented.
D000273:Adipose Tissue / Q000378:metabolism; D000345:Affinity Labels / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D001386:Azides / Q000378:metabolism; D001665:Binding Sites; D004591:Electrophoresis, Polyacrylamide Gel; D006207:Half-Life; D006863:Hydrogen-Ion Concentration; D007158:Immunologic Techniques; D007328:Insulin / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism*; D050356:Lipid Metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010777:Photochemistry; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D000345:Affinity Labels; D000906:Antibodies; D001386:Azides; D007328:Insulin; D010446:Peptide Fragments; C035459:insulin, (2-nitro-4-azidophenylacetyl)(B2)-des-Phe(B1)-; C036769:insulin, (2-nitro-4-azidophenylacetyl) (B29)-; D011972:Receptor, Insulin; D014357:Trypsin