Analysis of growth hormone and lactogenic binding sites cross-linked to iodinated human growth hormone.
GH (GHR) and lactogenic receptors were analyzed after use of the cross-linking reagent ethylene glycol bis-(succinimidyl succinate) to attach covalently iodinated human GH (hGH) to binding proteins 1) on intact IM-9 lymphocytes, 2) in a partially purified GHR preparation from rabbit liver, and 3) in crude microsomal fractions from rabbit liver, rabbit mammary gland, and rat liver. The latter two microsomal preparations contain primarily lactogenic receptors, whereas in IM-9 lymphocytes and the rabbit liver preparations, GHR predominate. Cross-linked [125I]hGH-receptor complexes were solubilized, reduced, and separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis of proteins cross-linked to [125I]hGH in the microsomal fraction from rabbit liver showed a specifically labeled complex with an estimated molecular weight (mol wt) of 75K. A slightly lower mol wt (71K) was determined for the complex labeled in the purified GHR preparation. In contrast to the relatively low mol wt complexes in rabbit liver, a complex that migrated with an apparent mol wt of 130K was identified in IM-9 lymphocytes. Labeled complexes were identified at 66K from rat liver and 61K from rabbit mammary gland. If it is assumed that hGH contributes 21K to the mol wt of the radiolabeled complexes, then the approximate mol wts of hGH-binding sites are 50-54K from rabbit liver, 109K from IM-9 lymphocytes, 45K from rat liver, and 40K from rabbit mammary gland.
D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D013006:Growth Hormone / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008099:Liver / Q000378:metabolism*; D008214:Lymphocytes / Q000378:metabolism; D046911:Macromolecular Substances; D008321:Mammary Glands, Animal / Q000378:metabolism; D008970:Molecular Weight; D010928:Placental Lactogen / Q000378:metabolism*; D011247:Pregnancy; D011817:Rabbits; D051381:Rats; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D018000:Receptors, Peptide; D011986:Receptors, Somatotropin
D007457:Iodine Radioisotopes; D046911:Macromolecular Substances; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; D011986:Receptors, Somatotropin; C028202:placental lactogen receptor; D013006:Growth Hormone; D010928:Placental Lactogen