Immunolocalization of sat-1 sulfate/oxalate/bicarbonate anion exchanger in the rat kidney.
The rat liver sulfate/bicarbonate/oxalate exchanger (sat-1) transports sulfate across the canalicular membrane in exchange for either bicarbonate or oxalate. Sulfate/oxalate exchange has been detected in the proximal tubule of the kidney, where it is probably involved in the reabsorption of filtered sulfate and the secretion of oxalate and may contribute to oxalate-dependent chloride reabsorption. Screening of a renal cortex cDNA library determined that sat-1 is expressed in the rat kidney. To evaluate this anion exchanger, the sat-1 protein was expressed in Sf9 cells. Sodium-independent sulfate and oxalate uptake was enhanced 7.3-fold and 13.1-fold, respectively, in Sf9 cells expressing the sat-1 protein compared with cells infected with wild-type virus. We determined that sat-1 is glycosylated in the kidney; however, anion exchange via sat-1 is observed despite incomplete glycosylation of sat-1 in Sf9 cells. The sat-1 protein, with an added COOH-terminal 6-histidine tag, was purified on a metal affinity column and used to generate anti-sat-1 monoclonal antibodies. The sat-1 protein was localized to the basolateral membrane, but not the apical membrane, of the proximal tubule by both Western blot analysis and immunohistochemistry. These studies demonstrate that sulfate/oxalate exchange on the apical and basolateral membranes of the proximal tubule represents transport on two different anion exchangers.
Laboratory of Epithelial Transport, Department of Internal Medicine, Department of Veterans Affairs Medical Center and University of Iowa, Iowa City, Iowa 52242, USA.
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LP
Karniski
M
M
Lötscher
M
M
Fucentese
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H
Hilfiker
J
J
Biber
H
H
Murer
D000818:Animals; D027321:Anion Transport Proteins; D017920:Antiporters / Q000032:analysis* / Q000096:biosynthesis*; D001639:Bicarbonates / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003001:Cloning, Molecular; D015723:Gene Library; D007150:Immunohistochemistry; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D007672:Kidney Cortex / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D010070:Oxalates / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D018411:Spodoptera; D000076988:Sulfate Transporters; D013431:Sulfates / Q000378:metabolism; D014162:Transfection
D027321:Anion Transport Proteins; D017920:Antiporters; D001639:Bicarbonates; D010070:Oxalates; D011994:Recombinant Proteins; C056459:Slc26a1 protein, rat; D000076988:Sulfate Transporters; D013431:Sulfates