Receptors for insulin and epidermal growth factor: interaction with organomercurial agarose.
The receptor for both insulin and epidermal growth factor (EGF) from human placental membranes, after crosslink labeling with 125I-labeled insulin and EGF, can be absorbed to an organomercurial-agarose derivative (Affi-Gel 501) and can be recovered from the gel by elution with dithiothreitol (DTT). Pretreatment of crosslink-labeled membranes with N-ethylmaleimide (NEM) blocks the ability of the receptor to react with the organomercurial column. NEM also abolishes the protein kinase activity of both receptors. Under appropriate conditions, insulin can promote the reaction of the insulin receptor with the organomercurial-agarose derivative. For both the insulin and EGF receptors, our results provide an avenue for the isolation of the sulfhydryl-containing receptor domains that may play a role in the control of receptor function.
D002846:Chromatography, Affinity; D004229:Dithiothreitol; D066246:ErbB Receptors; D005033:Ethylmaleimide; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology; D008565:Membrane Proteins / Q000302:isolation & purification; D009941:Organomercury Compounds; D010920:Placenta / Q000032:analysis; D011247:Pregnancy; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011972:Receptor, Insulin / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives*
D007328:Insulin; D008565:Membrane Proteins; D009941:Organomercury Compounds; D011956:Receptors, Cell Surface; C047528:Affi-Gel 501; D012685:Sepharose; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D005033:Ethylmaleimide; D004229:Dithiothreitol