Purification and immunological characterization of the rat liver insulin-like growth factor-II receptor.
Rat liver microsomal insulin-like growth factor-II (IGF-II) receptor has been purified to homogeneity using a single step affinity chromatographic procedure on agarose-IGF-II with elution at pH 4. Determined by either IGF-II binding or a direct RIA for receptor, purification of 2000-fold was obtained. The mean recovery was 28% for five such preparations. Sodium dodecyl sulfate-electrophoresis and autoradiography of purified receptor, radioiodinated receptor, and affinity-labeled receptor all indicated a molecular mass of approximately 250K. Scatchard analysis of IGF-II binding to purified receptor, solubilized microsomal membranes, or plasma membranes showed a single class of binding site with an affinity constant of 6 X 10(10) liter/mol in all cases. Potent antibodies to the purified receptor were raised in rabbits, capable of inhibiting 50% of IGF-II binding at dilutions of 1:170,000 and also of fully precipitating IGF-II-prelabeled receptor at 1:50,000. Both types of antibodies reacted with IGF-II receptors in rat adipose tissue, brain, heart, kidney, lung, and spleen. However, little cross-reactivity was seen with other species. Comparison of the ability of receptor antibodies to inhibit IGF-II binding to microsomal and plasma membranes indicated a specific immunological difference between the IGF-II receptors in the two membrane preparations.
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D003429:Cross Reactions; D008099:Liver / Q000502:physiology*; D008970:Molecular Weight; D051381:Rats; D011972:Receptor, Insulin / Q000276:immunology / Q000302:isolation & purification*; D017451:Receptors, Somatomedin; D013045:Species Specificity; D014018:Tissue Distribution
D017451:Receptors, Somatomedin; D011972:Receptor, Insulin