Localization and topology of a urate transporter/channel, a galectin, in epithelium-derived cells.
Recombinant protein produced from a cDNA cloned in our laboratory (UAT) functions in lipid bilayers as a urate transporter/channel. Because UAT is a galectin, a family of proteins presumed to be soluble, the localization and topology of UAT were assessed in living cells. UAT was targeted to plasma membrane in multiple epithelium-derived cell lines and, in polarized cells, was targeted to both apical and basolateral membranes. The amino and carboxy termini of UAT were both detected on the cytoplasmic side of plasma membranes, whereas cell surface biotinylation studies demonstrated that UAT is not merely a cytosolic membrane-associated protein but contains at least one extracellular domain. Madin-Darby canine kidney cells were shown both functionally and immunologically to contain an apparent homolog of UAT; however, transfection with UAT did not modify urate uptake. Because coimmunoprecipitation studies revealed that UAT is capable of forming both homo- and heteromultimers, it is proposed that monomers of endogenous channels are in part replaced by monomers of the protein expressed subsequent to transfection, thereby maintaining constancy of urate uptake at basal levels.
Rappoport
J Z
JZ
Division of Nephrology, Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029, USA.
Lipkowitz
M S
MS
Abramson
R G
RG
D000818:Animals; D019921:Biotinylation; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D037161:Galectins; D017930:Genes, Reporter; D006388:Hemagglutinins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D018613:Microscopy, Confocal; D027361:Organic Anion Transporters; D027701:Organic Cation Transport Proteins; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D014527:Uric Acid / Q000737:chemistry / Q000378:metabolism*
D002352:Carrier Proteins; D037161:Galectins; D006388:Hemagglutinins; D027361:Organic Anion Transporters; D027701:Organic Cation Transport Proteins; D011993:Recombinant Fusion Proteins; C491241:SLC22A12 protein, human; C103862:urate transporter; D014527:Uric Acid