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Title: Photoaffinity labeling of the human erythrocyte glucose transporter with /sup 4/H-labelled forskolin

Abstract

Forskolin, a potent activator of adenylate cyclase, is also known to inhibit glucose transport in a number of cells. The authors have investigated photoincorporation of (/sup 3/H)forskolin into erythrocyte membrane proteins using a technique they previously developed for photolabeling the erythrocyte glucose transporter with cytochalasin B (CB). A 30-40s irradiation of erythrocyte ghosts in the presence of (/sup 3/H)forskolin resulted in a concentration-dependent, covalent incorporation of radiolabel into all of the major membrane protein bands. However, most of the incorporation occurred in only three regions of the gel. Peak 1 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 (approx. 90kDa), and the third region labeled was between 41-60 kDa which corresponds to the region of the glucose transporter. This region appeared to contain several overlapping peaks with the largest incorporation of label occurring around 45 kDa in the area of red cell actin. When photolabeling was performed in the presence of 400 ..mu..M cytochalasin B (8.0 ..mu..M forskolin) the labeling in the 41-60 kDa region was totally inhibited while labeling of the 90 kDa peak was partially blocked. CB had no effect onmore » the photolabeling of peak 1 by forskolin.« less

Authors:
; ;
Publication Date:
Research Org.:
Southern Illinois Univ. School of Medicine, Carbondale
OSTI Identifier:
6898961
Report Number(s):
CONF-8606151-
Journal ID: CODEN: FEPRA; TRN: 87-007011
Resource Type:
Conference
Journal Name:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
Additional Journal Information:
Journal Volume: 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ENZYME INHIBITORS; BIOLOGICAL EFFECTS; GLUCOSE; MEMBRANE TRANSPORT; COVALENCE; ERYTHROCYTES; INHIBITION; LABELLING; MAN; PROTEINS; TRACER TECHNIQUES; TRITIUM COMPOUNDS; ALDEHYDES; ANIMALS; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CARBOHYDRATES; HEXOSES; ISOTOPE APPLICATIONS; LABELLED COMPOUNDS; MAMMALS; MATERIALS; MONOSACCHARIDES; ORGANIC COMPOUNDS; PRIMATES; SACCHARIDES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Shanahan, M F, Edwards, B M, and Morris, D P. Photoaffinity labeling of the human erythrocyte glucose transporter with /sup 4/H-labelled forskolin. United States: N. p., 1986. Web.
Shanahan, M F, Edwards, B M, & Morris, D P. Photoaffinity labeling of the human erythrocyte glucose transporter with /sup 4/H-labelled forskolin. United States.
Shanahan, M F, Edwards, B M, and Morris, D P. 1986. "Photoaffinity labeling of the human erythrocyte glucose transporter with /sup 4/H-labelled forskolin". United States.
@article{osti_6898961,
title = {Photoaffinity labeling of the human erythrocyte glucose transporter with /sup 4/H-labelled forskolin},
author = {Shanahan, M F and Edwards, B M and Morris, D P},
abstractNote = {Forskolin, a potent activator of adenylate cyclase, is also known to inhibit glucose transport in a number of cells. The authors have investigated photoincorporation of (/sup 3/H)forskolin into erythrocyte membrane proteins using a technique they previously developed for photolabeling the erythrocyte glucose transporter with cytochalasin B (CB). A 30-40s irradiation of erythrocyte ghosts in the presence of (/sup 3/H)forskolin resulted in a concentration-dependent, covalent incorporation of radiolabel into all of the major membrane protein bands. However, most of the incorporation occurred in only three regions of the gel. Peak 1 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 (approx. 90kDa), and the third region labeled was between 41-60 kDa which corresponds to the region of the glucose transporter. This region appeared to contain several overlapping peaks with the largest incorporation of label occurring around 45 kDa in the area of red cell actin. When photolabeling was performed in the presence of 400 ..mu..M cytochalasin B (8.0 ..mu..M forskolin) the labeling in the 41-60 kDa region was totally inhibited while labeling of the 90 kDa peak was partially blocked. CB had no effect on the photolabeling of peak 1 by forskolin.},
doi = {},
url = {https://www.osti.gov/biblio/6898961}, journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:6,
place = {United States},
year = {Thu May 01 00:00:00 EDT 1986},
month = {Thu May 01 00:00:00 EDT 1986}
}

Conference:
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