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Photolabeling of glucose-sensitive cytochalasin B binding proteins in erythrocyte, fibroblast and adipocyte membranes
Authors:M F Shanahan  S A Olson  M J Weber  G E Lienhard  J C Gorga
Institution:1. Department of Physiology, University of Wisconsin, Madison, WI 53706 USA;2. Department of Microbiology, University of Illinois, Urbana, IL 61801 USA;3. Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755 USA
Abstract:(3H)Cytochalasin B has been photoincorporated into membrane fractions of the human erythrocyte, Rous sarcoma virus-transformed chicken embryo fibroblast and rat adipocyte. Identification of D-glucose sensitive cytochalasin B binding sites was achieved by photolyzing membranes with radioligand in the presence of 0.5–0.7M D- or L-glucose. In the erythrocyte the major labeled bands on SDS-polyacrylamide gels were at 55,000 and 46,000 daltons. In the virus-transformed fibroblasts a major labeled band was at 55,000 daltons, and in adipocyte microsomal membranes, peaks at 50,000 and 45,000 daltons were observed. Binding characteristics of these polypeptides suggest that they are the putative glucose transport proteins in these three cell types.
Keywords:CB  cytochalasin B  CE  cytochalasin E  SDS-PAGE  sodium dodecyl sulfate gel electrophoresis
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