Reconstitution of the beef heart and rat liver mitochondrial K+/H+ (Na+/H+) antiporter. Quantitation of K+ transport with the novel fluorescent probe, PBFI
New indicators for fluorescent measurement of Na+ and K+ ions should prove particularly useful for studies of reconstituted carriers of these ions. We show that PBFI, a K(+)-specific probe, provides a convenient and sensitive assay for the study of K+ uptake mediated by the reconstituted mitochondri...
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| Veröffentlicht in: | The Journal of biological chemistry 1990-06, Vol.265 (18), p.10522-10526 |
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| container_title | The Journal of biological chemistry |
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| creator | JEZEK, P MAHDI, F GARLID, K. D |
| description | New indicators for fluorescent measurement of Na+ and K+ ions should prove particularly useful for studies of reconstituted
carriers of these ions. We show that PBFI, a K(+)-specific probe, provides a convenient and sensitive assay for the study
of K+ uptake mediated by the reconstituted mitochondrial K+/H+ (Na+/H+) antiporter. Fluorescent measurements have enabled
us for the first time to establish reconstitution of the K+/H+ (Na+/H+) antiporter from beef heart as well as from rat liver
mitochondria. This technique has also enabled us to establish that dicyclohexylcarbodiimide is capable of complete inhibition
of K+/H+ antiport in the reconstituted system, in accord with findings in intact mitochondria. PBFI fluorescence, which measures
net K+ uptake, was essential for this corroboration, since dicyclohexylcarbodiimide is not capable of complete inhibition
of 42K+/K+ or 86Rb+/Rb+ exchange, presumably because it acts selectively on proton transport within the carrier. |
| doi_str_mv | 10.1016/S0021-9258(18)86978-2 |
| format | Article |
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of K+ uptake mediated by the reconstituted mitochondrial K+/H+ (Na+/H+) antiporter. Fluorescent measurements have enabled
us for the first time to establish reconstitution of the K+/H+ (Na+/H+) antiporter from beef heart as well as from rat liver
mitochondria. This technique has also enabled us to establish that dicyclohexylcarbodiimide is capable of complete inhibition
of K+/H+ antiport in the reconstituted system, in accord with findings in intact mitochondria. PBFI fluorescence, which measures
net K+ uptake, was essential for this corroboration, since dicyclohexylcarbodiimide is not capable of complete inhibition
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carriers of these ions. We show that PBFI, a K(+)-specific probe, provides a convenient and sensitive assay for the study
of K+ uptake mediated by the reconstituted mitochondrial K+/H+ (Na+/H+) antiporter. Fluorescent measurements have enabled
us for the first time to establish reconstitution of the K+/H+ (Na+/H+) antiporter from beef heart as well as from rat liver
mitochondria. This technique has also enabled us to establish that dicyclohexylcarbodiimide is capable of complete inhibition
of K+/H+ antiport in the reconstituted system, in accord with findings in intact mitochondria. PBFI fluorescence, which measures
net K+ uptake, was essential for this corroboration, since dicyclohexylcarbodiimide is not capable of complete inhibition
of 42K+/K+ or 86Rb+/Rb+ exchange, presumably because it acts selectively on proton transport within the carrier.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Binding and carrier proteins</subject><subject>Biological and medical sciences</subject><subject>Biological Transport, Active</subject><subject>Carrier Proteins - metabolism</subject><subject>Fluorescent Dyes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Intracellular Membranes - metabolism</subject><subject>Kinetics</subject><subject>Liposomes</subject><subject>Mathematics</subject><subject>Mitochondria, Heart - enzymology</subject><subject>Mitochondria, Liver - enzymology</subject><subject>Potassium - metabolism</subject><subject>Potassium-Hydrogen Antiporters</subject><subject>Proteins</subject><subject>Proteolipids - metabolism</subject><subject>Sodium-Hydrogen Exchangers</subject><subject>Spectrometry, Fluorescence</subject><subject>Submitochondrial Particles - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFUdtu1DAUtBCoLIVPqOQHQK2WtLYTx_EjVJRWrbgj8WY5zjExSuLFdlrxLfwszu5S_HJkzZwZewahI0pOKaH12RdCGC0k480xbU6aWoqmYA_QipKmLEpOvz9Eq3vKY_Qkxp8kn0rSA3TAaM1Kzlboz2cwforJpTk5P2FvceoBtwAW96BDwnrqcNAJD-4WAh5d8qb3UxecHvD1-uxyjY_f62WeZGpyGx8ShFP8aV5uSf9TvV7jFPQUFxzfudRvfSZ_CwO2w-wDRANTwpvgW3iFP765uHqKHlk9RHi2n4fo28Xbr-eXxc2Hd1fnr28KU1GSCs5r2eZEKta0ksr8R4Ba1FBWnaWCiIa2phVSmtIaTS0QJkxtRMcq2_GWy_IQvdzpZu9fM8SkRpcfMwx6Aj9HJWRTZh2aiXxHNMHHGMCqTXCjDr8VJWopRW1LUUviijZqW4piee9obzC3I3T3W_sWMv5ij-to9GBzTsbF_-JSVELSMvOe73i9-9HfuQCqdbkNGBWr-WJICWes_AuybKB-</recordid><startdate>19900625</startdate><enddate>19900625</enddate><creator>JEZEK, P</creator><creator>MAHDI, F</creator><creator>GARLID, K. D</creator><general>American Society for Biochemistry and Molecular Biology</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>7X8</scope></search><sort><creationdate>19900625</creationdate><title>Reconstitution of the beef heart and rat liver mitochondrial K+/H+ (Na+/H+) antiporter. Quantitation of K+ transport with the novel fluorescent probe, PBFI</title><author>JEZEK, P ; MAHDI, F ; GARLID, K. D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c410t-5569b016428b919000ee676e34df170781bcb799c3fca1fe027c6c7d24fd5b593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Binding and carrier proteins</topic><topic>Biological and medical sciences</topic><topic>Biological Transport, Active</topic><topic>Carrier Proteins - metabolism</topic><topic>Fluorescent Dyes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Intracellular Membranes - metabolism</topic><topic>Kinetics</topic><topic>Liposomes</topic><topic>Mathematics</topic><topic>Mitochondria, Heart - enzymology</topic><topic>Mitochondria, Liver - enzymology</topic><topic>Potassium - metabolism</topic><topic>Potassium-Hydrogen Antiporters</topic><topic>Proteins</topic><topic>Proteolipids - metabolism</topic><topic>Sodium-Hydrogen Exchangers</topic><topic>Spectrometry, Fluorescence</topic><topic>Submitochondrial Particles - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>JEZEK, P</creatorcontrib><creatorcontrib>MAHDI, F</creatorcontrib><creatorcontrib>GARLID, K. D</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>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>JEZEK, P</au><au>MAHDI, F</au><au>GARLID, K. D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Reconstitution of the beef heart and rat liver mitochondrial K+/H+ (Na+/H+) antiporter. Quantitation of K+ transport with the novel fluorescent probe, PBFI</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1990-06-25</date><risdate>1990</risdate><volume>265</volume><issue>18</issue><spage>10522</spage><epage>10526</epage><pages>10522-10526</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>New indicators for fluorescent measurement of Na+ and K+ ions should prove particularly useful for studies of reconstituted
carriers of these ions. We show that PBFI, a K(+)-specific probe, provides a convenient and sensitive assay for the study
of K+ uptake mediated by the reconstituted mitochondrial K+/H+ (Na+/H+) antiporter. Fluorescent measurements have enabled
us for the first time to establish reconstitution of the K+/H+ (Na+/H+) antiporter from beef heart as well as from rat liver
mitochondria. This technique has also enabled us to establish that dicyclohexylcarbodiimide is capable of complete inhibition
of K+/H+ antiport in the reconstituted system, in accord with findings in intact mitochondria. PBFI fluorescence, which measures
net K+ uptake, was essential for this corroboration, since dicyclohexylcarbodiimide is not capable of complete inhibition
of 42K+/K+ or 86Rb+/Rb+ exchange, presumably because it acts selectively on proton transport within the carrier.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>2162352</pmid><doi>10.1016/S0021-9258(18)86978-2</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1990-06, Vol.265 (18), p.10522-10526 |
| issn | 0021-9258 1083-351X |
| language | eng |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Analytical, structural and metabolic biochemistry Binding and carrier proteins Biological and medical sciences Biological Transport, Active Carrier Proteins - metabolism Fluorescent Dyes Fundamental and applied biological sciences. Psychology Intracellular Membranes - metabolism Kinetics Liposomes Mathematics Mitochondria, Heart - enzymology Mitochondria, Liver - enzymology Potassium - metabolism Potassium-Hydrogen Antiporters Proteins Proteolipids - metabolism Sodium-Hydrogen Exchangers Spectrometry, Fluorescence Submitochondrial Particles - metabolism |
| title | Reconstitution of the beef heart and rat liver mitochondrial K+/H+ (Na+/H+) antiporter. Quantitation of K+ transport with the novel fluorescent probe, PBFI |
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