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Sodium-calcium exchange in renal epithelial cells : dependence on cell sodium and competitive inhibition by magnesium

Kinetic properties of Na(+)-Ca2+ exchange in a renal epithelial cell line (LLC-MK2) were assessed by measuring cytosolic free Ca2+ with fura-2 and 45Ca2+ influx. Replacing external Na+ with K+ produced relatively small increases in free Ca2+ and 45Ca2+ uptake unless the cells were incubated with oua...

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Published in:The Journal of membrane biology 1991-10, Vol.124 (1), p.73-83
Main Authors: RONG-MING LYU, SMITH, L, BINGHAM SMITH, J
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description Kinetic properties of Na(+)-Ca2+ exchange in a renal epithelial cell line (LLC-MK2) were assessed by measuring cytosolic free Ca2+ with fura-2 and 45Ca2+ influx. Replacing external Na+ with K+ produced relatively small increases in free Ca2+ and 45Ca2+ uptake unless the cells were incubated with ouabain. Ouabain markedly increased cell Na+ and strongly potentiated the effect of replacing external Na+ with K+ on free Ca2+ and 45Ca2+ uptake. 45Ca2+ influx in 140 mM K+ or N-methyl-D-glucamine minus influx in 140 mM Na+ was used to quantify Na(+)-Ca2+ exchange activity of Na(+)-loaded cells. The dependence of exchange on cell Na+ was sigmoidal; the K0.5 was 26 +/- 3 mmol/liter cell water space, and the Hill coefficient was 3.1 +/- 0.2. The kinetic features of the dependence of exchange on cell Na+ partly account for the small increase in Ca2+ influx when all external Na+ is replaced by K+. Besides raising cell Na+ ouabain appears to activate the exchanger. Magnesium competitively inhibited exchange activity. The potency of Mg2+ was 8.2-fold lower with potassium instead of N-methyl-D-glucamine or choline as the replacement for external Na+. Potassium also increased the Vmax of exchange by 86% and had no effect on the Km for Ca2+. The exchanger does not cause detectable 22Na(+)-Mg2+ exchange and does not appear to require K+ or transport 86Rb+. Although exchange activity was plentiful in the epithelial cells from monkey kidney, others from amphibian, canine, opossum, and porcine kidney had no detectable exchange activity. All of the measured kinetic properties of Na(+)-Ca2+ exchange in the renal epithelial cells are very similar to those of the exchanger in rat aortic myocytes.
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Replacing external Na+ with K+ produced relatively small increases in free Ca2+ and 45Ca2+ uptake unless the cells were incubated with ouabain. Ouabain markedly increased cell Na+ and strongly potentiated the effect of replacing external Na+ with K+ on free Ca2+ and 45Ca2+ uptake. 45Ca2+ influx in 140 mM K+ or N-methyl-D-glucamine minus influx in 140 mM Na+ was used to quantify Na(+)-Ca2+ exchange activity of Na(+)-loaded cells. The dependence of exchange on cell Na+ was sigmoidal; the K0.5 was 26 +/- 3 mmol/liter cell water space, and the Hill coefficient was 3.1 +/- 0.2. The kinetic features of the dependence of exchange on cell Na+ partly account for the small increase in Ca2+ influx when all external Na+ is replaced by K+. Besides raising cell Na+ ouabain appears to activate the exchanger. Magnesium competitively inhibited exchange activity. The potency of Mg2+ was 8.2-fold lower with potassium instead of N-methyl-D-glucamine or choline as the replacement for external Na+. 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Potassium also increased the Vmax of exchange by 86% and had no effect on the Km for Ca2+. The exchanger does not cause detectable 22Na(+)-Mg2+ exchange and does not appear to require K+ or transport 86Rb+. Although exchange activity was plentiful in the epithelial cells from monkey kidney, others from amphibian, canine, opossum, and porcine kidney had no detectable exchange activity. 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Psychology</subject><subject>Kidney - chemistry</subject><subject>Kidney - cytology</subject><subject>Kidney - physiology</subject><subject>Magnesium - pharmacology</subject><subject>Membrane and intracellular transports</subject><subject>Molecular and cellular biology</subject><subject>Ouabain - pharmacology</subject><subject>Potassium - pharmacology</subject><subject>sodium</subject><subject>Sodium - analysis</subject><subject>Sodium - pharmacology</subject><subject>Sodium - physiology</subject><subject>Sodium-Calcium Exchanger</subject><subject>Tetraethylammonium</subject><subject>Tetraethylammonium Compounds - pharmacology</subject><issn>0022-2631</issn><issn>1432-1424</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><recordid>eNqFkc1LxDAQxYMo6_px8S7kIB6EaibNJqk3FVcFwYN6Lmk6dSNtWptW9L83-4EePc0w78djZh4hR8DOgTF1cT1noBWkUm6RKYiUJyC42CZTxjhPuExhl-yF8M4YKCXFhExASq64mpLxuS3d2CTW1DZWil92YfwbUudpj97UFDs3LLB2sbVY14Fe0hI79CV6i7T1qykNKxtqfElt23Q4uMF9Ll0WrohtxIpv2pg3jyFyB2SnMnXAw03dJ6_z25eb--Tx6e7h5uoxsangQxK3rZhMuS60tqJiClAUAk0m40lCIWSFBVYiGtC8LDItSpVBVYHSwuj4iH1yuvbt-vZjxDDkjQvLfY3Hdgy54rNslmb8XxAkgGSZjODZGrR9G0KPVd71rjH9dw4sX4aR_4UR4eON61g0WP6h6-9H_WSjmxADqHrjrQu_2CympzWkP-hkkQI</recordid><startdate>19911001</startdate><enddate>19911001</enddate><creator>RONG-MING LYU</creator><creator>SMITH, L</creator><creator>BINGHAM SMITH, J</creator><general>Springer</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FD</scope><scope>FR3</scope><scope>M7Z</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19911001</creationdate><title>Sodium-calcium exchange in renal epithelial cells : dependence on cell sodium and competitive inhibition by magnesium</title><author>RONG-MING LYU ; SMITH, L ; BINGHAM SMITH, J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c342t-177f06328b88c4f071e4b4ea9663147e19bc10deea182db984d791ff1784a8143</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biological Transport - drug effects</topic><topic>Biological Transport - physiology</topic><topic>calcium</topic><topic>Calcium - pharmacokinetics</topic><topic>Calcium - pharmacology</topic><topic>Calcium Channels - drug effects</topic><topic>Calcium Channels - physiology</topic><topic>Carrier Proteins - physiology</topic><topic>Cell physiology</topic><topic>Cells, Cultured</topic><topic>Epithelial Cells</topic><topic>Epithelium - chemistry</topic><topic>Epithelium - physiology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Kidney - chemistry</topic><topic>Kidney - cytology</topic><topic>Kidney - physiology</topic><topic>Magnesium - pharmacology</topic><topic>Membrane and intracellular transports</topic><topic>Molecular and cellular biology</topic><topic>Ouabain - pharmacology</topic><topic>Potassium - pharmacology</topic><topic>sodium</topic><topic>Sodium - analysis</topic><topic>Sodium - pharmacology</topic><topic>Sodium - physiology</topic><topic>Sodium-Calcium Exchanger</topic><topic>Tetraethylammonium</topic><topic>Tetraethylammonium Compounds - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>RONG-MING LYU</creatorcontrib><creatorcontrib>SMITH, L</creatorcontrib><creatorcontrib>BINGHAM SMITH, J</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 1</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of membrane biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>RONG-MING LYU</au><au>SMITH, L</au><au>BINGHAM SMITH, J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sodium-calcium exchange in renal epithelial cells : dependence on cell sodium and competitive inhibition by magnesium</atitle><jtitle>The Journal of membrane biology</jtitle><addtitle>J Membr Biol</addtitle><date>1991-10-01</date><risdate>1991</risdate><volume>124</volume><issue>1</issue><spage>73</spage><epage>83</epage><pages>73-83</pages><issn>0022-2631</issn><eissn>1432-1424</eissn><coden>JMBBBO</coden><abstract>Kinetic properties of Na(+)-Ca2+ exchange in a renal epithelial cell line (LLC-MK2) were assessed by measuring cytosolic free Ca2+ with fura-2 and 45Ca2+ influx. Replacing external Na+ with K+ produced relatively small increases in free Ca2+ and 45Ca2+ uptake unless the cells were incubated with ouabain. Ouabain markedly increased cell Na+ and strongly potentiated the effect of replacing external Na+ with K+ on free Ca2+ and 45Ca2+ uptake. 45Ca2+ influx in 140 mM K+ or N-methyl-D-glucamine minus influx in 140 mM Na+ was used to quantify Na(+)-Ca2+ exchange activity of Na(+)-loaded cells. The dependence of exchange on cell Na+ was sigmoidal; the K0.5 was 26 +/- 3 mmol/liter cell water space, and the Hill coefficient was 3.1 +/- 0.2. The kinetic features of the dependence of exchange on cell Na+ partly account for the small increase in Ca2+ influx when all external Na+ is replaced by K+. Besides raising cell Na+ ouabain appears to activate the exchanger. Magnesium competitively inhibited exchange activity. The potency of Mg2+ was 8.2-fold lower with potassium instead of N-methyl-D-glucamine or choline as the replacement for external Na+. Potassium also increased the Vmax of exchange by 86% and had no effect on the Km for Ca2+. The exchanger does not cause detectable 22Na(+)-Mg2+ exchange and does not appear to require K+ or transport 86Rb+. Although exchange activity was plentiful in the epithelial cells from monkey kidney, others from amphibian, canine, opossum, and porcine kidney had no detectable exchange activity. All of the measured kinetic properties of Na(+)-Ca2+ exchange in the renal epithelial cells are very similar to those of the exchanger in rat aortic myocytes.</abstract><cop>New York, NY</cop><pub>Springer</pub><pmid>1662727</pmid><doi>10.1007/BF01871366</doi><tpages>11</tpages></addata></record>
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ispartof The Journal of membrane biology, 1991-10, Vol.124 (1), p.73-83
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subjects Animals
Biological and medical sciences
Biological Transport - drug effects
Biological Transport - physiology
calcium
Calcium - pharmacokinetics
Calcium - pharmacology
Calcium Channels - drug effects
Calcium Channels - physiology
Carrier Proteins - physiology
Cell physiology
Cells, Cultured
Epithelial Cells
Epithelium - chemistry
Epithelium - physiology
Fundamental and applied biological sciences. Psychology
Kidney - chemistry
Kidney - cytology
Kidney - physiology
Magnesium - pharmacology
Membrane and intracellular transports
Molecular and cellular biology
Ouabain - pharmacology
Potassium - pharmacology
sodium
Sodium - analysis
Sodium - pharmacology
Sodium - physiology
Sodium-Calcium Exchanger
Tetraethylammonium
Tetraethylammonium Compounds - pharmacology
title Sodium-calcium exchange in renal epithelial cells : dependence on cell sodium and competitive inhibition by magnesium
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