skip to main content

Title: Characterization of mammalian glucose transport proteins using photoaffinity labeling techniques

A carrier-free radioiodinated phenylazide derivative of forskolin, 3-iodo-4-azidophenethylamido-7-O-succinyl-deacetyl-forskolin (({sup 125}I)IAPS-forskolin), has been shown to be a highly selective photoaffinity probe for the human erythrocyte glucose transported and the glucose transport proteins found in several mammalian tissues and cultured cells where the glucose transport protein is present at a low concentration. The photoincorporation of ({sup 125}I)IAPS-forskolin into these glucose transporters was blocked by D- (but not L-) glucose, cytochalasin B, and forskolin. In addition to labeling the mammalian glucose transport proteins, ({sup 125}I)IAPS-forskolin also labeled the L-arabinose transporter from E. coli. In muscle and adipose tissues, glucose transport is markedly increased in response to insulin. ({sup 125}I)IAPS-forskolin was shown to selectivity tag the glucose transporter in membranes derived from these cells. In addition, the covalent derivatization of the transport protein in subcellular fractions of the adipocyte has provided a means to study the hormonal regulation of glucose transport. ({sup 125}I)IAPS-forskolin has also been used to label the purified human erythrocyte glucose transporter. The site of insertion has therefore been localized by analysis of the radiolabeled peptides which were produced following chemical and proteolytic digestion of the labeled transport protein.
Authors:
Publication Date:
OSTI Identifier:
6275092
Resource Type:
Miscellaneous
Resource Relation:
Other Information: Thesis (Ph. D.)
Publisher:
Madison, WI (US); Univ. of Wisconsin
Research Org:
Wisconsin Univ., Madison, WI (USA)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; GLUCOSE; MEMBRANE TRANSPORT; MEMBRANE PROTEINS; CHEMICAL COMPOSITION; ADIPOSE TISSUE; CELL MEMBRANES; ERYTHROCYTES; INSULIN; IODINE 125; LABELLING; MAN; MUSCLES; PHOSPHOLIPIDS; TRACER TECHNIQUES; ALDEHYDES; ANIMAL TISSUES; ANIMALS; BETA DECAY RADIOISOTOPES; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY; BODY FLUIDS; CARBOHYDRATES; CELL CONSTITUENTS; CONNECTIVE TISSUE; DAYS LIVING RADIOISOTOPES; ELECTRON CAPTURE RADIOISOTOPES; ESTERS; HEXOSES; HORMONES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; LIPIDS; MAMMALS; MATERIALS; MEMBRANES; MONOSACCHARIDES; NUCLEI; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; ORGANIC PHOSPHORUS COMPOUNDS; PEPTIDE HORMONES; PRIMATES; PROTEINS; RADIOISOTOPES; SACCHARIDES; TISSUES; VERTEBRATES 550501* -- Metabolism-- Tracer Techniques