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Am J Physiol Renal Physiol. 2017 Aug 1;313(2):F467-F474. doi: 10.1152/ajprenal.00628.2016. Epub 2017 Jun 7.

Characterization of the transport activity of SGLT2/MAP17, the renal low-affinity Na+-glucose cotransporter.

Author information

1
Groupe d'Étude des Protéines Membranaires, Département de Physique, Université de Montréal, Montréal, Québec, Canada.
2
Groupe d'Étude des Protéines Membranaires, Département de Physique, Université de Montréal, Montréal, Québec, Canada jean-yves.lapointe@umontreal.ca.

Abstract

The cotransporter SGLT2 is responsible for 90% of renal glucose reabsorption, and we recently showed that MAP17 appears to work as a required β-subunit. We report in the present study a detailed functional characterization of human SGLT2 in coexpression with human MAP17 in Xenopus laevis oocytes. Addition of external glucose generates a large inward current in the presence of Na, confirming an electrogenic transport mechanism. At a membrane potential of -50 mV, SGLT2 affinity constants for glucose and Na are 3.4 ± 0.4 and 18 ± 6 mM, respectively. The change in the reversal potential of the cotransport current as a function of external glucose concentration clearly confirms a 1:1 Na-to-glucose transport stoichiometry. SGLT2 is selective for glucose and α-methylglucose but also transports, to a lesser extent, galactose and 3-O-methylglucose. SGLT2 can be inhibited in a competitive manner by phlorizin (Ki = 31 ± 4 nM) and by dapagliflozin (Ki = 0.75 ± 0.3 nM). Similarly to SGLT1, SGLT2 can be activated by Na, Li, and protons. Pre-steady-state currents for SGLT2 do exist but are small in amplitude and relatively fast (a time constant of ~2 ms). The leak current defined as the phlorizin-sensitive current in the absence of substrate was extremely small in the case of SGLT2. In summary, in comparison with SGLT1, SGLT2 has a lower affinity for glucose, a transport stoichiometry of 1:1, very small pre-steady-state and leak currents, a 10-fold higher affinity for phlorizin, and an affinity for dapagliflozin in the subnanomolar range.

KEYWORDS:

dapagliflozin; electrophysiology; glucose transport; kinetics; transport properties

Comment in

PMID:
28592437
DOI:
10.1152/ajprenal.00628.2016
[Indexed for MEDLINE]
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