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PMID: 3047124 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Studies with antipeptide antibody suggest the presence of at least two types of glucose transporter in rat brain and adipocyte.

The Journal of biological chemistry ·Vol. 263 ·No. 26 ·1988-09-15 ·Pages 13432-9

Oka Y, Asano T, Shibasaki Y, Kasuga M, Kanazawa Y, Takaku F

Abstract

Three antipeptide antibodies were prepared by immunizing rabbits with synthesized short peptides corresponding to residues 215-226, 466-479, and 478-492 predicted from the cDNA of both the human hepatoma HepG2 and rat brain glucose transporters. All three antibodies were found to precipitate quantitatively the [3H]cytochalasin B photoaffinity-labeled human erythrocyte glucose transporter. Each antibody also recognized the rat brain protein of Mr 45,000 on immunoblots, and a similar molecular weight protein was labeled with [3H]cytochalasin B in a D-glucose-inhibitable manner, suggesting that this protein is glucose transporter. However, only up to 30% of the labeled rat brain glucose transporters were precipitated, even by repeated rounds of immunoprecipitation. In addition, these antibodies were observed to be unable to immunoprecipitate significantly the [3H]cytochalasin B-labeled rat adipocyte glucose transporter. Further, one-dimensional peptide maps of [3H]cytochalasin B-labeled human erythrocyte and adipocyte glucose transporters generated distinct tryptic fragments. Although Mr 45,000 protein in rat adipocyte low density microsomes was detected on immunoblots and its amount was decreased in insulin-treated cells, the rat adipocyte low density microsomes were much less reactive on immunoblots than the rat brain membranes in spite of the fact that the rat adipocyte low density microsomes contained more [3H]cytochalasin B-labeled glucose transporters. In addition, the ratio of cytochalasin B-labeled glucose transporter per unit HepG2-type glucose transporter mRNA was more than 10-fold higher in rat adipocyte than in rat brain. These results indicate that virtually all the human erythrocyte glucose transporters are of the HepG2 type, whereas this type of glucose transporter constitutes only approximately 30 and 3% of all the glucose transporters present in rat brain and rat adipocyte, respectively; and the rest, of similar molecular weight, is expressed by a different gene.

MeSH Terms
Adipose Tissue/analysis Affinity Labels/metabolism Animals Antibodies Brain Chemistry Erythrocytes/analysis Humans Immunosorbent Techniques Liver Neoplasms, Experimental/analysis Monosaccharide Transport Proteins/analysis Photochemistry Rats
Chemicals
Affinity Labels Antibodies Monosaccharide Transport Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Oka Y
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Asano T
Shibasaki Y
Kasuga M
Kanazawa Y
Takaku F
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
Published
1988-09-15
Pages
13432-9
Language
English
Region
United States
NLM ID
2985121R
Subset
IM
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