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Reconstitution, identification, purification, and immunological characterization of the 110-kDa Na+/Ca2+ antiporter from beef heart mitochondria
The mitochondrial Na+/Ca2+ antiporter plays a key role in the physiological regulation of intramitochondrial Ca2+, which in turn attunes mitochondrial enzymes to the changing demands of the cell for ATP. We have now purified the Na+/Ca2+ antiporter from beef heart mitochondria by assaying detergent-...
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| Published in: | The Journal of biological chemistry 1992-09, Vol.267 (25), p.17983-17989 |
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| Main Authors: | , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c430t-97bbe7fdc915b820a921dc4c1b82f94b932ea2fe4b142dff7f8f273b818a37823 |
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| cites | cdi_FETCH-LOGICAL-c430t-97bbe7fdc915b820a921dc4c1b82f94b932ea2fe4b142dff7f8f273b818a37823 |
| container_end_page | 17989 |
| container_issue | 25 |
| container_start_page | 17983 |
| container_title | The Journal of biological chemistry |
| container_volume | 267 |
| creator | Li, W Shariat-Madar, Z Powers, M Sun, X Lane, R D Garlid, K D |
| description | The mitochondrial Na+/Ca2+ antiporter plays a key role in the physiological regulation of intramitochondrial Ca2+, which in
turn attunes mitochondrial enzymes to the changing demands of the cell for ATP. We have now purified the Na+/Ca2+ antiporter
from beef heart mitochondria by assaying detergent-solubilized chromatography fractions for reconstitutive activity. Na+ and
Ca2+ transport were assayed using the fluorescent probes, sodium-binding benzofuran isophthalate and Fura-2, respectively.
This approach enabled us to identify Na+/Ca2+ exchange activity with a 110-kDa inner membrane protein that catalyzed Na(+)-dependent
Ca2+ transport and Ca(2+)-dependent Na+ transport. A new finding was that the Na+/Ca2+ antiporter also catalyzed Na+/Li+ exchange
in the absence of Ca2+. All modes of transport were electroneutral and were inhibited by diltiazem and tetraphenylphosphonium
cation. Monospecific polyclonal antibodies to the 110-kDa protein inhibited Na+/Ca2+ and Na+/Li+ exchange in the reconstituted
system and recognized 110-kDa proteins in mitochondrial membranes isolated from rat heart, liver, and kidney. |
| doi_str_mv | 10.1016/s0021-9258(19)37140-6 |
| format | article |
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turn attunes mitochondrial enzymes to the changing demands of the cell for ATP. We have now purified the Na+/Ca2+ antiporter
from beef heart mitochondria by assaying detergent-solubilized chromatography fractions for reconstitutive activity. Na+ and
Ca2+ transport were assayed using the fluorescent probes, sodium-binding benzofuran isophthalate and Fura-2, respectively.
This approach enabled us to identify Na+/Ca2+ exchange activity with a 110-kDa inner membrane protein that catalyzed Na(+)-dependent
Ca2+ transport and Ca(2+)-dependent Na+ transport. A new finding was that the Na+/Ca2+ antiporter also catalyzed Na+/Li+ exchange
in the absence of Ca2+. All modes of transport were electroneutral and were inhibited by diltiazem and tetraphenylphosphonium
cation. Monospecific polyclonal antibodies to the 110-kDa protein inhibited Na+/Ca2+ and Na+/Li+ exchange in the reconstituted
system and recognized 110-kDa proteins in mitochondrial membranes isolated from rat heart, liver, and kidney.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/s0021-9258(19)37140-6</identifier><identifier>PMID: 1517231</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>Animals ; Calcium - metabolism ; Carrier Proteins - isolation & purification ; Carrier Proteins - metabolism ; Cattle ; Chromatography, DEAE-Cellulose ; Chromatography, Gel ; Diltiazem - pharmacology ; Electrophoresis, Polyacrylamide Gel ; Intracellular Membranes - metabolism ; Kinetics ; Mitochondria, Heart - metabolism ; Models, Biological ; Molecular Weight ; Sodium - metabolism ; Sodium-Calcium Exchanger ; Time Factors</subject><ispartof>The Journal of biological chemistry, 1992-09, Vol.267 (25), p.17983-17989</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c430t-97bbe7fdc915b820a921dc4c1b82f94b932ea2fe4b142dff7f8f273b818a37823</citedby><cites>FETCH-LOGICAL-c430t-97bbe7fdc915b820a921dc4c1b82f94b932ea2fe4b142dff7f8f273b818a37823</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1517231$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, W</creatorcontrib><creatorcontrib>Shariat-Madar, Z</creatorcontrib><creatorcontrib>Powers, M</creatorcontrib><creatorcontrib>Sun, X</creatorcontrib><creatorcontrib>Lane, R D</creatorcontrib><creatorcontrib>Garlid, K D</creatorcontrib><title>Reconstitution, identification, purification, and immunological characterization of the 110-kDa Na+/Ca2+ antiporter from beef heart mitochondria</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The mitochondrial Na+/Ca2+ antiporter plays a key role in the physiological regulation of intramitochondrial Ca2+, which in
turn attunes mitochondrial enzymes to the changing demands of the cell for ATP. We have now purified the Na+/Ca2+ antiporter
from beef heart mitochondria by assaying detergent-solubilized chromatography fractions for reconstitutive activity. Na+ and
Ca2+ transport were assayed using the fluorescent probes, sodium-binding benzofuran isophthalate and Fura-2, respectively.
This approach enabled us to identify Na+/Ca2+ exchange activity with a 110-kDa inner membrane protein that catalyzed Na(+)-dependent
Ca2+ transport and Ca(2+)-dependent Na+ transport. A new finding was that the Na+/Ca2+ antiporter also catalyzed Na+/Li+ exchange
in the absence of Ca2+. All modes of transport were electroneutral and were inhibited by diltiazem and tetraphenylphosphonium
cation. Monospecific polyclonal antibodies to the 110-kDa protein inhibited Na+/Ca2+ and Na+/Li+ exchange in the reconstituted
system and recognized 110-kDa proteins in mitochondrial membranes isolated from rat heart, liver, and kidney.</description><subject>Animals</subject><subject>Calcium - metabolism</subject><subject>Carrier Proteins - isolation & purification</subject><subject>Carrier Proteins - metabolism</subject><subject>Cattle</subject><subject>Chromatography, DEAE-Cellulose</subject><subject>Chromatography, Gel</subject><subject>Diltiazem - pharmacology</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Intracellular Membranes - metabolism</subject><subject>Kinetics</subject><subject>Mitochondria, Heart - metabolism</subject><subject>Models, Biological</subject><subject>Molecular Weight</subject><subject>Sodium - metabolism</subject><subject>Sodium-Calcium Exchanger</subject><subject>Time Factors</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><recordid>eNpNkd9qFTEQxoMo9Xj0EQq5EFHq2kyyu9lcyrFqoSj4B7wLSTbpRnc3xySLtE_hIzenW6xzM3yZ75vAbxA6BvIGCLSniRAKlaBN9xLEK8ahJlX7AG2AdKxiDfx4iDb_LI_Rk5R-klK1gCN0BA1wymCD_n6xJswp-7xkH-bX2Pd2zt55o1a9X-J_Ss099tO0zGEMl-V1xGZQUZlso7--9eDgcB4sBiDVr3cKf1InpztFT0o0-32IxYldDBPW1jo8WBUznnwOZghzH716ih45NSb77K5v0ff3Z992H6uLzx_Od28vKlMzkivBtbbc9UZAoztKlKDQm9pAEU7UWjBqFXW21lDT3jnuOkc50x10ivGOsi16se7dx_B7sSnLySdjx1HNNixJcgYt8MJyi5rVaGJIKVon99FPKl5JIPJwCfn1gFkeMEsQ8vYSsi2547sPFj3Z_j61oi_z5-t88JfDHx-t1L5QsJOkLZe0kcBFx9gNo1-STA</recordid><startdate>19920905</startdate><enddate>19920905</enddate><creator>Li, W</creator><creator>Shariat-Madar, Z</creator><creator>Powers, M</creator><creator>Sun, X</creator><creator>Lane, R D</creator><creator>Garlid, K D</creator><general>American Society for Biochemistry and Molecular Biology</general><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>19920905</creationdate><title>Reconstitution, identification, purification, and immunological characterization of the 110-kDa Na+/Ca2+ antiporter from beef heart mitochondria</title><author>Li, W ; Shariat-Madar, Z ; Powers, M ; Sun, X ; Lane, R D ; Garlid, K D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c430t-97bbe7fdc915b820a921dc4c1b82f94b932ea2fe4b142dff7f8f273b818a37823</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Animals</topic><topic>Calcium - metabolism</topic><topic>Carrier Proteins - isolation & purification</topic><topic>Carrier Proteins - metabolism</topic><topic>Cattle</topic><topic>Chromatography, DEAE-Cellulose</topic><topic>Chromatography, Gel</topic><topic>Diltiazem - pharmacology</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Intracellular Membranes - metabolism</topic><topic>Kinetics</topic><topic>Mitochondria, Heart - metabolism</topic><topic>Models, Biological</topic><topic>Molecular Weight</topic><topic>Sodium - metabolism</topic><topic>Sodium-Calcium Exchanger</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, W</creatorcontrib><creatorcontrib>Shariat-Madar, Z</creatorcontrib><creatorcontrib>Powers, M</creatorcontrib><creatorcontrib>Sun, X</creatorcontrib><creatorcontrib>Lane, R D</creatorcontrib><creatorcontrib>Garlid, K D</creatorcontrib><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>Li, W</au><au>Shariat-Madar, Z</au><au>Powers, M</au><au>Sun, X</au><au>Lane, R D</au><au>Garlid, K D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Reconstitution, identification, purification, and immunological characterization of the 110-kDa Na+/Ca2+ antiporter from beef heart mitochondria</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1992-09-05</date><risdate>1992</risdate><volume>267</volume><issue>25</issue><spage>17983</spage><epage>17989</epage><pages>17983-17989</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The mitochondrial Na+/Ca2+ antiporter plays a key role in the physiological regulation of intramitochondrial Ca2+, which in
turn attunes mitochondrial enzymes to the changing demands of the cell for ATP. We have now purified the Na+/Ca2+ antiporter
from beef heart mitochondria by assaying detergent-solubilized chromatography fractions for reconstitutive activity. Na+ and
Ca2+ transport were assayed using the fluorescent probes, sodium-binding benzofuran isophthalate and Fura-2, respectively.
This approach enabled us to identify Na+/Ca2+ exchange activity with a 110-kDa inner membrane protein that catalyzed Na(+)-dependent
Ca2+ transport and Ca(2+)-dependent Na+ transport. A new finding was that the Na+/Ca2+ antiporter also catalyzed Na+/Li+ exchange
in the absence of Ca2+. All modes of transport were electroneutral and were inhibited by diltiazem and tetraphenylphosphonium
cation. Monospecific polyclonal antibodies to the 110-kDa protein inhibited Na+/Ca2+ and Na+/Li+ exchange in the reconstituted
system and recognized 110-kDa proteins in mitochondrial membranes isolated from rat heart, liver, and kidney.</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>1517231</pmid><doi>10.1016/s0021-9258(19)37140-6</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1992-09, Vol.267 (25), p.17983-17989 |
| issn | 0021-9258 1083-351X |
| language | eng |
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| source | ScienceDirect |
| subjects | Animals Calcium - metabolism Carrier Proteins - isolation & purification Carrier Proteins - metabolism Cattle Chromatography, DEAE-Cellulose Chromatography, Gel Diltiazem - pharmacology Electrophoresis, Polyacrylamide Gel Intracellular Membranes - metabolism Kinetics Mitochondria, Heart - metabolism Models, Biological Molecular Weight Sodium - metabolism Sodium-Calcium Exchanger Time Factors |
| title | Reconstitution, identification, purification, and immunological characterization of the 110-kDa Na+/Ca2+ antiporter from beef heart mitochondria |
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