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/sup 3/H)forskolin. Direct photoaffinity labeling of the erythrocyte D-glucose transporter

Journal Article · · J. Biol. Chem.; (United States)
OSTI ID:6498762
; ;

Irradiation of erythrocyte ghosts in the presence of (/sup 3/H)forskolin resulted in a concentration-dependent, covalent incorporation of radiolabel into several of the major membrane protein bands. Most of the incorporation occurred in four regions of the gel. Peak 1 (216 kDa) was a sharp peak near the top of the gel in the region corresponding to spectrin. Peak 2 appeared to be associated with band 3 (89 kDa), while a third peak occurred around the position of band 4.2 (76 kDa). The fourth region of labeling was a broad area between 43-75 kDa which corresponds to the region of the glucose transporter. Forskolin labeling of this region was inhibited by cytochalasin B and D-glucose, but not L-glucose. Extraction of extrinsic membrane proteins resulted in a loss of radiolabeled protein from the 216- and 76-kDa regions. Treatment of membranes labeled with either cytochalasin B or forskolin with endo-beta-galactosidase resulted in identical shifts of the 43 to 75-kDa peaks to 42 kDa. Similarly, trypsinization of membranes photolabeled with either cytochalasin B or forskolin resulted in the generation of a 17-kDa radiolabeled fragment in both cases. Photoincorporation of (/sup 3/H)cytochalasin B into the glucose transporter was blocked in a concentration-dependent manner by unlabeled forskolin.

Research Organization:
Southern Illinois Univ. School of Medicine, Carbondale
OSTI ID:
6498762
Journal Information:
J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 13; ISSN JBCHA
Country of Publication:
United States
Language:
English

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Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALDEHYDES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
CHEMISTRY
DOSE-RESPONSE RELATIONSHIPS
ERYTHROCYTES
GLUCOSE
HEXOSES
IRRADIATION
LABELLED COMPOUNDS
LABELLING
MATERIALS
MEMBRANE TRANSPORT
MEMBRANES
MOLECULAR WEIGHT
MONOSACCHARIDES
ORGANIC COMPOUNDS
PHOTOCHEMISTRY
PROTEINS
SACCHARIDES
TRITIUM COMPOUNDS