Expression of a rat renal sodium-dependent dicarboxylate transporter in Xenopus oocytes.
Microinjection of mRNA isolated from rat kidney cortex into Xenopus laevis oocytes resulted in the expression of a Na(+)-dependent dicarboxylate transporter, as detected by uptake measurements with [14C]succinate as substrate. The expressed transporter showed an S-shaped Na(+)-dependence with half-maximal activation at 19-21 mM Na+ and a Hill coefficient between 2 and 3. Endogenous succinate uptake was not Na(+)-dependent. Na(+)-stimulated succinate uptake in mRNA-injected oocytes exhibited a maximum at pH 7.5, whereas endogenous Na(+)-independent transporter was fastest at pH 8.5. The expressed dicarboxylate transporter also differed from the endogenous transporter in its sensitivity to citrate as well as dicarboxylates in trans and cis configurations. The expressed transporter resembled the renal basolateral transporter, especially with respect to affinity for succinate (Km 28 microM), activation by Na+, pH-dependence and substrate specificity. After injection of size-fractionated mRNA, succinate uptake was expressed by mRNA of 2-3 kb. Our results suggest expression of the basolateral Na(+)-dependent dicarboxylate transporter after injection of mRNA from rat kidney into Xenopus oocytes.
Steffgen
J
J
Sektion Nephrologie, Universität Ulm, Germany.
Kienle
S
S
Scheyerl
F
F
Franz
H E
HE
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D029382:Dicarboxylic Acid Transporters; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006863:Hydrogen-Ion Concentration; D007672:Kidney Cortex / Q000737:chemistry*; D008845:Microinjections; D009865:Oocytes / Q000378:metabolism*; D012333:RNA, Messenger / Q000008:administration & dosage / Q000378:metabolism; D051381:Rats; D012964:Sodium / Q000494:pharmacology*; D013386:Succinates / Q000378:metabolism; D019802:Succinic Acid; D014982:Xenopus laevis
D002352:Carrier Proteins; D029382:Dicarboxylic Acid Transporters; D012333:RNA, Messenger; D013386:Succinates; D012964:Sodium; D019802:Succinic Acid