SAAT1 is a low affinity Na+/glucose cotransporter and not an amino acid transporter. A reinterpretation.
Recently a member of the Na+/glucose (SGLT1) gene family of cotransporters was isolated from a pig renal cell line and was thought to be the neutral amino acid transporter System A. This cDNA (Kong, C. T., Yet, S. F., and Lever, J. E. (1993) J. Biol. Chem. 268, 1509-1512) encodes a 660-amino acid protein with 76% identity to SGLT1. To confirm and extend the kinetic characterization of SAAT1, we have expressed this clone in Xenopus oocytes and measured transport using both radiotracer and electrophysiological techniques. SAAT1 did not stimulate either 50 microM 2-(methylamino)isobutyrate uptake or 2-(methylamino)isobutyrate-evoked inward Na+ currents, but instead stimulated 50 microM alpha MG (alpha-methyl-D-glucopyranoside) uptake 27-fold from 2 +/- 1 pmol.h-1/oocyte (n = 9) to 55 +/- 6 pmol.h-1/oocyte (n = 9) and alpha MG-evoked inward Na+ currents (I) by up to 1000 nA/oocyte. The apparent affinity constant for alpha MG (K alpha MG 0.5) was approximately 2 mM and was independent of membrane potential from -30 to -150 mV but was voltage-sensitive between -30 and +30 mV. The relative sugar specificity for the transporter was alpha MG > or = D-glucose >> D-galactose >>> 3-O-methyl-D-glucopyranose, L-glucose. The sugar-evoked currents were Na(+)-dependent (KNa 0.5 approximately 10 mM at -50 mV) and the Hill coefficient was 1. KNa 0.5 decreased with hyperpolarization of the membrane from -50 to -150 mV. Phlorizin inhibited the alpha MG-evoked current with apparent Ki of 18 microM at -50 mV. We conclude that the SAAT1 cDNA encodes a renal low affinity Na+(1)/glucose(1) cotransporter and propose that pig SAAT1 be renamed pSGLT2.
Mackenzie
B
B
Department of Physiology, UCLA School of Medicine 90024.
Panayotova-Heiermann
M
M
Loo
D D
DD
Lever
J E
JE
Wright
E M
EM
D026905:Amino Acid Transport Systems; D000621:Aminoisobutyric Acids / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D002250:Carbon Radioisotopes; D002352:Carrier Proteins / Q000096:biosynthesis / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D005071:Evoked Potentials / Q000187:drug effects; D005260:Female; D015870:Gene Expression; D005947:Glucose / Q000031:analogs & derivatives* / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis / Q000378:metabolism*; D009865:Oocytes / Q000502:physiology*; D013379:Substrate Specificity; D013552:Swine; D014982:Xenopus laevis
D026905:Amino Acid Transport Systems; D000621:Aminoisobutyric Acids; D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; C017911:2-(methylamino)isobutyric acid; D005947:Glucose