rapid insulin-dependent endocytosis of the insulin receptor by caveolae in primary adipocytes快速胰岛素依赖型小窝胰岛素受体的内吞作用的主要组织.pdfVIP

Rapid Insulin-Dependent Endocytosis of the Insulin Receptor by Caveolae in Primary Adipocytes ¨ ˚ Siri Fagerholm, Unn Ortegren, Margareta Karlsson, Iida Ruishalme, Peter Stralfors* ¨ ¨ Division of Cell Biology, Department of Clinical and Experimental Medicine and Diabetes Research Centre, University of Linkoping, Linkoping, Sweden Abstract Background: The insulin receptor is localized in caveolae and is dependent on caveolae or cholesterol for signaling in adipocytes. When stimulated with insulin, the receptor is internalized. Methodology/Principal Findings: We examined primary rat adipocytes by subcellular fractionation to examine if the insulin receptor was internalized in a caveolae-mediated process. Insulin induced a rapid, t1/2,3 min, endocytosis of the insulin receptor in parallel with receptor tyrosine autophosphorylation. Concomitantly, caveolin-1 was phosphorylated at tyrosine(14) and endocytosed. Vanadate increased the phosphorylation of caveolin-1 without affecting insulin receptor phosphorylation or endocytosis. Immunocapture of endosomal vesicles with antibodies against the insulin receptor co- captured caveolin-1 and immunocapture with antibodies against tyrosine(14)-phosphorylated caveolin-1 co-captured the insulin receptor, demonstrating that the insulin receptor was endocytosed together with tyrosine(14)-phosphorylated caveolin-1. By immun