Cloning and functional characterization of the human GLUT7 isoform SLC2A7 from the small intestine.
Facilitated glucose transporters (GLUTs) mediate transport of sugars across cell membranes by using the chemical gradient of sugars as the driving force. Improved cloning techniques and database analyses have expanded this family of proteins to a total of 14 putative members. In this work a novel hexose transporter isoform, GLUT7, has been cloned from a human intestinal cDNA library by using a PCR-based strategy (GenBank accession no. AY571960). The encoded protein is comprised of 524 amino acid residues and shares 68% similarity and 53% identity with GLUT5, its most closely related isoform. When GLUT7 was expressed in Xenopus oocytes, it showed high-affinity transport for glucose (K(m) = 0.3 mM) and fructose (IC(50) = 0.060 mM). Galactose, 2-deoxy-d-glucose, and xylose were not transported. Uptake of 100 microM d-glucose was not inhibited by 200 microM phloretin or 100 microM cytochalasin B. Northern blotting indicated that the mRNA for GLUT7 is present in the human small intestine, colon, testis, and prostate. Western blotting and immunohistochemistry of rat tissues with an antibody raised against the predicted COOH-terminal sequence confirmed expression of the protein in the small intestine and indicated that the transporter is predominantly expressed in the enterocytes' brush-border membrane. The unusual substrate specificity and close sequence identity with GLUT5 suggest that GLUT7 represents an intermediate between class II GLUTs and the class I member GLUT2. Comparison between these proteins may provide key information as to the structural determinants for the recognition of fructose as a substrate.
Li
Qiang
Q
Department of Physiology, University of Alberta, Rm. 7-22 Medical Sciences Bldg., Edmonton, Alberta T6G 2H7, Canada.
Manolescu
Andrei
A
Ritzel
Mabel
M
Yao
Sylvia
S
Slugoski
Melissa
M
Young
James D
JD
Chen
Xing-Zhen
XZ
Cheeseman
Chris I
CI
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D003001:Cloning, Molecular; D003106:Colon / Q000378:metabolism; D003571:Cytochalasin B / Q000494:pharmacology; D005260:Female; D005632:Fructose / Q000493:pharmacokinetics; D005947:Glucose / Q000493:pharmacokinetics; D051246:Glucose Transport Proteins, Facilitative; D006801:Humans; D007421:Intestine, Small / Q000378:metabolism*; D008297:Male; D008871:Microvilli / Q000378:metabolism; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D009865:Oocytes; D010693:Phloretin / Q000494:pharmacology; D011467:Prostate / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000378:metabolism; D014018:Tissue Distribution; D014982:Xenopus laevis
D051246:Glucose Transport Proteins, Facilitative; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; C487892:SLC2A7 protein, human; D005632:Fructose; D003571:Cytochalasin B; D005947:Glucose; D010693:Phloretin