Interaction of H + with the extracellular and intracellular aspects of hMATE1

Yodying Dangprapai, Stephen Wright

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Human multidrug and toxin extrusion 1 (hMATE1, SLC47A1) is a major candidate for being the molecular identity of organic cation/proton (OC/H +) exchange activity in the luminal membrane of renal proximal tubules. Although physiological function of hMATE1 supports luminal OC efflux, the kinetics of hMATE1-mediated OC transport have typically been characterized through measurement of uptake, i.e., the interaction between outward-facing hMATE1 and OCs. To examine kinetics of hMATE1-mediated transport in a more physiologically relevant direction, i.e., an interaction between inward-facing hMATE1 and cytoplasmic substrates, we measured the time course of hMATE1- mediated efflux of the prototypic MATE1 substrate, [3H]1-methyl-4- phenylpyridinium, under a variety of intra- and extracellular pH conditions, from Chinese hamster ovary cells that stably expressed the transporter. In this study, we showed that an IC50/Ki for interaction between extracellular H + and outward-facing hMATE1 determined from conventional uptake experiments [12.9 ± 1.23 nM (pH 7.89); n = 9] and from the efflux protocol [14.7 ± 3.45 nM (pH 7.83); n = 3] was not significantly different (P = 0.6). Furthermore, kinetics of interaction between intracellular H_ and inward-facing hMATE1 determined using the efflux protocol revealed an IC50 for H + of 11.5 nM (pH 7.91), consistent with symmetrical interactions of H + with the inward-facing and outward-facing aspects of hMATE1.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume301
Issue number3
DOIs
StatePublished - Sep 2011

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Inhibitory Concentration 50
1-Methyl-4-phenylpyridinium
Proximal Kidney Tubule
Cricetulus
Cations
Protons
Ovary
Membranes
Direction compound

Keywords

  • Organic cation
  • pH
  • Renal proximal tubule
  • Secretion
  • Transport

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

Interaction of H + with the extracellular and intracellular aspects of hMATE1. / Dangprapai, Yodying; Wright, Stephen.

In: American Journal of Physiology - Renal Physiology, Vol. 301, No. 3, 09.2011.

Research output: Contribution to journalArticle

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