Cloning and functional expression of a human pancreatic islet glucose-transporter cDNA.
Previous studies have suggested that pancreatic islet glucose transport is mediated by a high-Km, low-affinity facilitated transporter similar to that expressed in liver. To determine the relationship between islet and liver glucose transporters, liver-type glucose-transporter cDNA clones were isolated from a human liver cDNA library. The liver-type glucose-transporter cDNA clone hybridized to mRNA transcripts of the same size in human liver and pancreatic islet RNA. A cDNA library was prepared from purified human pancreatic islet tissue and screened with human liver-type glucose-transporter cDNA. We isolated two overlapping cDNA clones encompassing 2600 base pairs, which encode a pancreatic islet protein identical in sequence to that of the putative liver-type glucose-transporter protein. Xenopus oocytes injected with synthetic mRNA transcribed from a full-length cDNA construct exhibited increased uptake of 2-deoxyglucose, confirming the functional identity of the clone. These cDNA clones can now be used to study regulation of expression of the gene and to assess the role of inherited defects in this gene as a candidate for inherited susceptibility to non-insulin-dependent diabetes mellitus.
Permutt
M A
MA
Department of Internal Medicine, Washington University School of Medicine, Saint Louis, MO 63110.
Koranyi
L
L
Keller
K
K
Lacy
P E
PE
Scharp
D W
DW
Mueckler
M
M
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D015723:Gene Library; D005796:Genes; D006801:Humans; D007515:Islets of Langerhans / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000235:genetics*; D009865:Oocytes / Q000378:metabolism; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D014981:Xenopus
D009004:Monosaccharide Transport Proteins; D012313:RNA; D004247:DNA