title: Binding of insulin-like growth Factor ii and multiplication-stimulating activity-stimulated phosphorylation in basolateral membranes from dog kidney.
abstract: Biologic actions of insulin and insulin-like growth factors (IGFs) are thought to be initiated by binding of peptides to tissues, followed by phosphorylation of specific hormone receptors. Both insulin and IGF bind to renal membranes, suggesting functional roles for these peptides in kidney. The present studies further characterize the interaction of multiplication-stimulating activity (MSA)/IGF II with its renal receptor. Specific binding of 125I-IGF II was measured in basolateral membranes isolated from proximal tubular cells of dog kidney. Binding was half-maximal at 10(-9) M MSA and was not inhibited by human growth hormone, IGF I, insulin, or anti-insulin receptor antibodies. Concentration-dependent MSA-stimulated phosphorylation of a Mr 135,000 protein band was demonstrated in autoradiograms of sodium dodecyl sulfate-polyacrylamide gels from basolateral membrane suspensions. Insulin increased phosphorylation of this band only in the presence of MSA, while a Mr 92,000 band was consistently phosphorylated with insulin alone. The phosphorylated Mr 135,000 band which had been solubilized with detergent from basolateral membranes was immunoprecipitated using serum from a patient with anti-insulin receptor antibodies suggesting that the band is the alpha subunit of the insulin receptor. This was supported by the demonstration of covalent cross-linkage of 125I-insulin to the Mr 135,000 band. We conclude that receptor-mediated MSA-stimulated phosphorylation of isolated basolateral membranes may reflect a process by which biological actions of IGF II are mediated in vivo. Our data suggest that insulin and IGF II may interact by regulating protein phosphorylation.
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D006133:Growth Substances / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology*; D007335:Insulin-Like Growth Factor II; D007668:Kidney / Q000378:metabolism*; D007672:Kidney Cortex / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology*; D010766:Phosphorylation; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000378:metabolism*
chemical_list: D006133:Growth Substances; D007328:Insulin; D010455:Peptides; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D013002:Somatomedins; D007335:Insulin-Like Growth Factor II