Phosphate transport by the human renal cotransporter NaPi-3 expressed in HEK-293 cells

The American Journal of Physiology
R T Timmer, Robert B Gunn

Abstract

The human renal Na-PO4 cotransporter gene NaPi-3 was expressed in human embryonic kidney HEK-293 cells, and the transport characteristics were measured in cells transfected with a vector containing NaPi-3 or with the vector alone (sham transfected). The initial rate of 32PO4 influx had saturation kinetics for external Na and PO4 with K1/2Na of 128 mM (PO4 = 0.1 mM) and K1/2PO4 of 0.084 mM (extracellular Na = 143 mM) in sham- and NaPi-3-transfected cells expressing the transporter. Transfection had no effect on the Na-independent 32PO4 influx, but transfection increased Na-dependent 32PO4 influxes 2.5- to 5-fold. Of the alkali cations, only Na significantly supported PO4 influx. Arsenate inhibited flux with an inhibition constant of 0.4 mM. The phosphate transport in sham- and NaPi-3-transfected cells has nearly the same temperature dependence in the absence and presence of extracellular Na. The Na-dependent phosphate flux decreased with pH in sham-transfected cells but was pH independent in transfected cells. The Na-dependent 32PO4 influx was inhibited by p-chloromercuriphenylsulfonate, phosphonoformate, phloretin, vanadate, and 5-(N-methyl-N-isobutyl)-amiloride but not by amiloride or other amiloride analogs. These functional ...Continue Reading

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Citations

Sep 24, 1999·Current Opinion in Nephrology and Hypertension·H S Tenenhouse
Apr 4, 2003·American Journal of Physiology. Cell Physiology·Sammy Elmariah, Robert B Gunn
Jan 7, 2005·The Journal of Pharmacology and Experimental Therapeutics·Odile SergentDominique Lagadic-Gossmann
Nov 6, 2009·Genes & Nutrition·Blythe D ShepardPamela L Tuma
Jul 22, 2018·Cell and Tissue Research·Sadaf N KalamChristopher R Murphy
Aug 30, 2000·The Journal of General Physiology·R T Timmer, R B Gunn
May 18, 2006·American Journal of Physiology. Renal Physiology·Balasubramaniem AshokkumarHamid M Said
Apr 13, 2004·American Journal of Physiology. Renal Physiology·Brian E PeerceRebecca D Clarke
Jun 6, 2000·American Journal of Physiology. Cell Physiology·X J MengR F Abercrombie
Dec 17, 2004·American Journal of Physiology. Cell Physiology·Mikiko ItoKen-ichi Miyamoto
Oct 15, 2013·Cell Stress & Chaperones·Melinda E TóthMiklós Sántha
Oct 4, 2000·Physiological Reviews·H MurerJ Biber
Oct 28, 2006·Alcoholism, Clinical and Experimental Research·Benita L McVickerCarol A Casey
Oct 12, 1999·The American Journal of Physiology·I FernandesC Silve
Jan 14, 1999·The American Journal of Physiology·R T Timmer, R B Gunn
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Apr 23, 2005·Toxicology and Applied Pharmacology·Christy C Bridges, Rudolfs K Zalups
Jul 7, 2021·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Marasarakottige Y ThanujaSangly P Srinivas

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