Single copies of subunits d, oligomycin-sensitivity conferring protein, and b are present in the Saccharomyces cerevisiae mitochondrial ATP synthase
In the mitochondrial ATP synthase (mtATPase) of the yeast Saccharomyces cerevisiae, the stoichiometry of subunits d, oligomycin-sensitivity conferring protein (OSCP), and b is poorly defined. We have investigated the stoichiometry of these subunits by the application of hexahistidine affinity purifi...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 1999-03, Vol.274 (11), p.7462-7466 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 7466 |
|---|---|
| container_issue | 11 |
| container_start_page | 7462 |
| container_title | The Journal of biological chemistry |
| container_volume | 274 |
| creator | Bateson, M Devenish, R.J Nagley, P Prescott, M |
| description | In the mitochondrial ATP synthase (mtATPase) of the yeast Saccharomyces cerevisiae, the stoichiometry of subunits d, oligomycin-sensitivity conferring protein (OSCP), and b is poorly defined. We have investigated the stoichiometry of these subunits by the application of hexahistidine affinity purification technology. We have previously demonstrated that intact mtATPase complexes incorporating a Hex6-tagged subunit can be isolated via Ni2+-nitrilotriacetic acid affinity chromatography (Bateson, M., Devenish, R. J., Nagley, P., and Prescott, M. (1996) Anal. Biochem. 238, 14-18). Strains were constructed in which Hex6-tagged versions of subunits d, OSCP, and b were coexpressed with the corresponding wild-type subunit. This coexpression resulted in a mixed population of mtATPase complexes containing untagged wild-type and Hex6-tagged subunits. The stoichiometry of each subunit was then assessed by determining whether or not the untagged wild-type subunit could be recovered from Ni2+-nitrilotriacetic acid purifications as an integral component of those complexes absorbed by virtue of the Hex6-tagged subunit. As only the Hex6-tagged subunit was recovered from such purifications, we demonstrate that the stoichiometry of subunits d, OSCP, and b in yeast is 1 in each case. |
| doi_str_mv | 10.1074/jbc.274.11.7462 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_69606034</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>69606034</sourcerecordid><originalsourceid>FETCH-LOGICAL-f262t-218d016f7dc3ea39d2cdcd140f897039215c82cca68e8236802f8700357cc323</originalsourceid><addsrcrecordid>eNqF0E2LFDEQBuAgijuunr1pTp62x6qkJ0kfl2X9gAWFGcFbk0mqZ7J0p8ckvTD_wx9sZNezuRSEp96iirG3CGsE3X6837u10O0aca1bJZ6xFYKRjdzgz-dsBSCw6cTGXLBXOd9DfW2HL9kFAihlEFfs9zbEw0jczadAmc8Dz8t-iaFk7q_4PIbDPJ1diE2mmEMJD6GcK44DpVQ7-SnNhUK84jZ6vuc2Uf2iigsPkZcj8a117mjT35g6wFGih5CDJT6FMrvjHH0KduTXu-88n2M52kyv2YvBjpnePNVLtvt0u7v50tx9-_z15vquGYQSpRFoPKAatHeSrOy8cN55bGEwnQbZCdw4I5yzypARUhkQg9EAcqOdk0Jesg-PsXWJXwvl0k8hOxpHG2lecq86BQpk-1-IGrXpUFX47gku-4l8f0phsunc_7t3Be8fwWDn3h5SyP2PrQCUILqaolr5B8Ftjc4</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17178916</pqid></control><display><type>article</type><title>Single copies of subunits d, oligomycin-sensitivity conferring protein, and b are present in the Saccharomyces cerevisiae mitochondrial ATP synthase</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Bateson, M ; Devenish, R.J ; Nagley, P ; Prescott, M</creator><creatorcontrib>Bateson, M ; Devenish, R.J ; Nagley, P ; Prescott, M</creatorcontrib><description>In the mitochondrial ATP synthase (mtATPase) of the yeast Saccharomyces cerevisiae, the stoichiometry of subunits d, oligomycin-sensitivity conferring protein (OSCP), and b is poorly defined. We have investigated the stoichiometry of these subunits by the application of hexahistidine affinity purification technology. We have previously demonstrated that intact mtATPase complexes incorporating a Hex6-tagged subunit can be isolated via Ni2+-nitrilotriacetic acid affinity chromatography (Bateson, M., Devenish, R. J., Nagley, P., and Prescott, M. (1996) Anal. Biochem. 238, 14-18). Strains were constructed in which Hex6-tagged versions of subunits d, OSCP, and b were coexpressed with the corresponding wild-type subunit. This coexpression resulted in a mixed population of mtATPase complexes containing untagged wild-type and Hex6-tagged subunits. The stoichiometry of each subunit was then assessed by determining whether or not the untagged wild-type subunit could be recovered from Ni2+-nitrilotriacetic acid purifications as an integral component of those complexes absorbed by virtue of the Hex6-tagged subunit. As only the Hex6-tagged subunit was recovered from such purifications, we demonstrate that the stoichiometry of subunits d, OSCP, and b in yeast is 1 in each case.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.274.11.7462</identifier><identifier>PMID: 10066811</identifier><language>eng</language><publisher>United States</publisher><subject>Adenosine Triphosphatases - chemistry ; Adenosine Triphosphatases - metabolism ; binding ; Carrier Proteins ; chromatography ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; h+-transporting atp synthase ; immobilized metal ion affinity chromatography ; Membrane Proteins - chemistry ; Membrane Proteins - metabolism ; metal ions ; mitochondria ; Mitochondria - enzymology ; Mitochondrial Proton-Translocating ATPases ; nickel ; nitrilotriacetic acid ; protein subunits ; proteins ; Proton-Translocating ATPases - chemistry ; Proton-Translocating ATPases - isolation & purification ; Proton-Translocating ATPases - metabolism ; purification ; pyrophosphatases ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - enzymology ; stoichiometry</subject><ispartof>The Journal of biological chemistry, 1999-03, Vol.274 (11), p.7462-7466</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10066811$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bateson, M</creatorcontrib><creatorcontrib>Devenish, R.J</creatorcontrib><creatorcontrib>Nagley, P</creatorcontrib><creatorcontrib>Prescott, M</creatorcontrib><title>Single copies of subunits d, oligomycin-sensitivity conferring protein, and b are present in the Saccharomyces cerevisiae mitochondrial ATP synthase</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>In the mitochondrial ATP synthase (mtATPase) of the yeast Saccharomyces cerevisiae, the stoichiometry of subunits d, oligomycin-sensitivity conferring protein (OSCP), and b is poorly defined. We have investigated the stoichiometry of these subunits by the application of hexahistidine affinity purification technology. We have previously demonstrated that intact mtATPase complexes incorporating a Hex6-tagged subunit can be isolated via Ni2+-nitrilotriacetic acid affinity chromatography (Bateson, M., Devenish, R. J., Nagley, P., and Prescott, M. (1996) Anal. Biochem. 238, 14-18). Strains were constructed in which Hex6-tagged versions of subunits d, OSCP, and b were coexpressed with the corresponding wild-type subunit. This coexpression resulted in a mixed population of mtATPase complexes containing untagged wild-type and Hex6-tagged subunits. The stoichiometry of each subunit was then assessed by determining whether or not the untagged wild-type subunit could be recovered from Ni2+-nitrilotriacetic acid purifications as an integral component of those complexes absorbed by virtue of the Hex6-tagged subunit. As only the Hex6-tagged subunit was recovered from such purifications, we demonstrate that the stoichiometry of subunits d, OSCP, and b in yeast is 1 in each case.</description><subject>Adenosine Triphosphatases - chemistry</subject><subject>Adenosine Triphosphatases - metabolism</subject><subject>binding</subject><subject>Carrier Proteins</subject><subject>chromatography</subject><subject>Chromatography, Affinity</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>h+-transporting atp synthase</subject><subject>immobilized metal ion affinity chromatography</subject><subject>Membrane Proteins - chemistry</subject><subject>Membrane Proteins - metabolism</subject><subject>metal ions</subject><subject>mitochondria</subject><subject>Mitochondria - enzymology</subject><subject>Mitochondrial Proton-Translocating ATPases</subject><subject>nickel</subject><subject>nitrilotriacetic acid</subject><subject>protein subunits</subject><subject>proteins</subject><subject>Proton-Translocating ATPases - chemistry</subject><subject>Proton-Translocating ATPases - isolation & purification</subject><subject>Proton-Translocating ATPases - metabolism</subject><subject>purification</subject><subject>pyrophosphatases</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - enzymology</subject><subject>stoichiometry</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0E2LFDEQBuAgijuunr1pTp62x6qkJ0kfl2X9gAWFGcFbk0mqZ7J0p8ckvTD_wx9sZNezuRSEp96iirG3CGsE3X6837u10O0aca1bJZ6xFYKRjdzgz-dsBSCw6cTGXLBXOd9DfW2HL9kFAihlEFfs9zbEw0jczadAmc8Dz8t-iaFk7q_4PIbDPJ1diE2mmEMJD6GcK44DpVQ7-SnNhUK84jZ6vuc2Uf2iigsPkZcj8a117mjT35g6wFGih5CDJT6FMrvjHH0KduTXu-88n2M52kyv2YvBjpnePNVLtvt0u7v50tx9-_z15vquGYQSpRFoPKAatHeSrOy8cN55bGEwnQbZCdw4I5yzypARUhkQg9EAcqOdk0Jesg-PsXWJXwvl0k8hOxpHG2lecq86BQpk-1-IGrXpUFX47gku-4l8f0phsunc_7t3Be8fwWDn3h5SyP2PrQCUILqaolr5B8Ftjc4</recordid><startdate>19990312</startdate><enddate>19990312</enddate><creator>Bateson, M</creator><creator>Devenish, R.J</creator><creator>Nagley, P</creator><creator>Prescott, M</creator><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>19990312</creationdate><title>Single copies of subunits d, oligomycin-sensitivity conferring protein, and b are present in the Saccharomyces cerevisiae mitochondrial ATP synthase</title><author>Bateson, M ; Devenish, R.J ; Nagley, P ; Prescott, M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f262t-218d016f7dc3ea39d2cdcd140f897039215c82cca68e8236802f8700357cc323</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Adenosine Triphosphatases - chemistry</topic><topic>Adenosine Triphosphatases - metabolism</topic><topic>binding</topic><topic>Carrier Proteins</topic><topic>chromatography</topic><topic>Chromatography, Affinity</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>h+-transporting atp synthase</topic><topic>immobilized metal ion affinity chromatography</topic><topic>Membrane Proteins - chemistry</topic><topic>Membrane Proteins - metabolism</topic><topic>metal ions</topic><topic>mitochondria</topic><topic>Mitochondria - enzymology</topic><topic>Mitochondrial Proton-Translocating ATPases</topic><topic>nickel</topic><topic>nitrilotriacetic acid</topic><topic>protein subunits</topic><topic>proteins</topic><topic>Proton-Translocating ATPases - chemistry</topic><topic>Proton-Translocating ATPases - isolation & purification</topic><topic>Proton-Translocating ATPases - metabolism</topic><topic>purification</topic><topic>pyrophosphatases</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - enzymology</topic><topic>stoichiometry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bateson, M</creatorcontrib><creatorcontrib>Devenish, R.J</creatorcontrib><creatorcontrib>Nagley, P</creatorcontrib><creatorcontrib>Prescott, M</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</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>Bateson, M</au><au>Devenish, R.J</au><au>Nagley, P</au><au>Prescott, M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Single copies of subunits d, oligomycin-sensitivity conferring protein, and b are present in the Saccharomyces cerevisiae mitochondrial ATP synthase</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1999-03-12</date><risdate>1999</risdate><volume>274</volume><issue>11</issue><spage>7462</spage><epage>7466</epage><pages>7462-7466</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>In the mitochondrial ATP synthase (mtATPase) of the yeast Saccharomyces cerevisiae, the stoichiometry of subunits d, oligomycin-sensitivity conferring protein (OSCP), and b is poorly defined. We have investigated the stoichiometry of these subunits by the application of hexahistidine affinity purification technology. We have previously demonstrated that intact mtATPase complexes incorporating a Hex6-tagged subunit can be isolated via Ni2+-nitrilotriacetic acid affinity chromatography (Bateson, M., Devenish, R. J., Nagley, P., and Prescott, M. (1996) Anal. Biochem. 238, 14-18). Strains were constructed in which Hex6-tagged versions of subunits d, OSCP, and b were coexpressed with the corresponding wild-type subunit. This coexpression resulted in a mixed population of mtATPase complexes containing untagged wild-type and Hex6-tagged subunits. The stoichiometry of each subunit was then assessed by determining whether or not the untagged wild-type subunit could be recovered from Ni2+-nitrilotriacetic acid purifications as an integral component of those complexes absorbed by virtue of the Hex6-tagged subunit. As only the Hex6-tagged subunit was recovered from such purifications, we demonstrate that the stoichiometry of subunits d, OSCP, and b in yeast is 1 in each case.</abstract><cop>United States</cop><pmid>10066811</pmid><doi>10.1074/jbc.274.11.7462</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 1999-03, Vol.274 (11), p.7462-7466 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_69606034 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Adenosine Triphosphatases - chemistry Adenosine Triphosphatases - metabolism binding Carrier Proteins chromatography Chromatography, Affinity Electrophoresis, Polyacrylamide Gel h+-transporting atp synthase immobilized metal ion affinity chromatography Membrane Proteins - chemistry Membrane Proteins - metabolism metal ions mitochondria Mitochondria - enzymology Mitochondrial Proton-Translocating ATPases nickel nitrilotriacetic acid protein subunits proteins Proton-Translocating ATPases - chemistry Proton-Translocating ATPases - isolation & purification Proton-Translocating ATPases - metabolism purification pyrophosphatases Saccharomyces cerevisiae Saccharomyces cerevisiae - enzymology stoichiometry |
| title | Single copies of subunits d, oligomycin-sensitivity conferring protein, and b are present in the Saccharomyces cerevisiae mitochondrial ATP synthase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-11T14%3A56%3A47IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Single%20copies%20of%20subunits%20d,%20oligomycin-sensitivity%20conferring%20protein,%20and%20b%20are%20present%20in%20the%20Saccharomyces%20cerevisiae%20mitochondrial%20ATP%20synthase&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Bateson,%20M&rft.date=1999-03-12&rft.volume=274&rft.issue=11&rft.spage=7462&rft.epage=7466&rft.pages=7462-7466&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.274.11.7462&rft_dat=%3Cproquest_pubme%3E69606034%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17178916&rft_id=info:pmid/10066811&rfr_iscdi=true |