Selective solubilization of membrane proteins differentially labeled by p-chloromercuribenzenesulfonic acid in the presence of sucrose
Broadbean (Vicia faba L.) leaf discs have been incubated with the slowly permeant thiol reagent [203Hg]-para-chloromercuribenzenesulfonic acid (PCMBS) in the presence or in the absence of sucrose, and the release of PCMBS-labeled proteins has been monitored in media containing various concentrations...
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| Veröffentlicht in: | Plant Physiol.; (United States) 1987-03, Vol.83 (3), p.541-545 |
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| creator | M'Batchi, B Pichelin, D Delrot, S |
| description | Broadbean (Vicia faba L.) leaf discs have been incubated with the slowly permeant thiol reagent [203Hg]-para-chloromercuribenzenesulfonic acid (PCMBS) in the presence or in the absence of sucrose, and the release of PCMBS-labeled proteins has been monitored in media containing various concentrations of urea, ethyleneglycol-bis-(β-aminoethyl ether)-N, N, N′, N′-tetraacetic acid (EGTA), sodium cholate, sodium dodecyl sulfate, Triton X-100, octylglucoside or (3-[3-cholamidopropyl)-dimethylammonio] 1-propane-sulfonate) (CHAPS). The proteins differentially labeled by PCMBS in the presence of sucrose which, on the basis of previous results, are assumed to include the sucrose carrier, were preferentially solubilized by 1% CHAPS, 1% octylglucoside, or 1% Triton X-100. Other PCMBS-labeled proteins ('background' proteins) could be partially removed by EGTA, urea, or 0.1% cholate. Sequential treatment by 10 mM EGTA and 1% CHAPS was found to give a fraction highly enriched in the differentially labeled proteins. Analysis of the specific activity of microsomal pellets suggests that the results obtained with leaf discs give a good account of what is occurring at the plasma membrane level. These data, which suggest that the proteins differentially labeled by PCMBS in the presence of sucrose are intrinsic membrane proteins, can be used to solubilize these proteins from microsomal fractions. |
| doi_str_mv | 10.1104/pp.83.3.541 |
| format | Article |
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The proteins differentially labeled by PCMBS in the presence of sucrose which, on the basis of previous results, are assumed to include the sucrose carrier, were preferentially solubilized by 1% CHAPS, 1% octylglucoside, or 1% Triton X-100. Other PCMBS-labeled proteins ('background' proteins) could be partially removed by EGTA, urea, or 0.1% cholate. Sequential treatment by 10 mM EGTA and 1% CHAPS was found to give a fraction highly enriched in the differentially labeled proteins. Analysis of the specific activity of microsomal pellets suggests that the results obtained with leaf discs give a good account of what is occurring at the plasma membrane level. These data, which suggest that the proteins differentially labeled by PCMBS in the presence of sucrose are intrinsic membrane proteins, can be used to solubilize these proteins from microsomal fractions.</description><identifier>ISSN: 0032-0889</identifier><identifier>EISSN: 1532-2548</identifier><identifier>DOI: 10.1104/pp.83.3.541</identifier><identifier>PMID: 16665285</identifier><identifier>CODEN: PPHYA5</identifier><language>eng</language><publisher>Rockville, MD: American Society of Plant Physiologists</publisher><subject>550201 - Biochemistry- Tracer Techniques ; Absorption. Translocation of ions and substances. Permeability ; ADDITIVES ; Agronomy. Soil science and plant productions ; BACTERIA ; BASIC BIOLOGICAL SCIENCES ; BETA DECAY RADIOISOTOPES ; BETA-MINUS DECAY RADIOISOTOPES ; Biological and medical sciences ; CARBOHYDRATES ; CELL CONSTITUENTS ; CELL MEMBRANES ; Cell physiology ; DAYS LIVING RADIOISOTOPES ; DETERGENTS ; DIFFERENTIAL SOLUBILIZATION ; DISACCHARIDES ; Economic plant physiology ; EMULSIFIERS ; ETIQUETADO ; EVEN-ODD NUCLEI ; FEUILLE ; Fundamental and applied biological sciences. Psychology ; Gels ; HEAVY NUCLEI ; HOJAS ; ISOTOPE APPLICATIONS ; ISOTOPES ; LABELLING ; LEAVES ; LEGUMINOSAE ; MARQUAGE ; Membrane and intracellular transports ; Membrane proteins ; MEMBRANE TRANSPORT ; MEMBRANES ; Membranes and Bioenergetics ; MERCURY 203 ; MERCURY ISOTOPES ; MICROORGANISMS ; Molecular and cellular biology ; NUCLEI ; Nutrition. Photosynthesis. Respiration. Metabolism ; OLIGOSACCHARIDES ; ORGANIC ACIDS ; ORGANIC COMPOUNDS ; ORGANIC SULFUR COMPOUNDS ; PLANTS ; PROTEINAS ; PROTEINE ; PROTEINS ; Radioactive decay ; RADIOISOTOPES ; Reagents ; RESPONSE MODIFYING FACTORS ; RHIZOBIUM ; SACCHARIDES ; SACCHAROSE ; Sodium ; SOLUBILIDAD ; SOLUBILITE ; SOLUBILITY ; Solubilization ; SUCROSA ; SUCROSE ; Sugars ; SULFONIC ACIDS ; SURFACE ACTIVE AGENTS ; SURFACTANT ; SURFACTANTES ; SURFACTANTS ; THIOL ; THIOLS ; TIOLES ; TRACER TECHNIQUES ; VICIA ; VICIA FABA ; WETTING AGENTS</subject><ispartof>Plant Physiol.; (United States), 1987-03, Vol.83 (3), p.541-545</ispartof><rights>Copyright 1987 American Society of Plant Physiologists</rights><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3581-7f5d4d22c13f35ebc7483352ba27345ef393a23aa783a255c31284e775542bb13</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/4270463$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/4270463$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,780,784,803,885,27923,27924,58016,58249</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7433279$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16665285$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/6479506$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>M'Batchi, B</creatorcontrib><creatorcontrib>Pichelin, D</creatorcontrib><creatorcontrib>Delrot, S</creatorcontrib><creatorcontrib>Centre National de la Recherche Scientifique (UA 574), Poitiers, France</creatorcontrib><title>Selective solubilization of membrane proteins differentially labeled by p-chloromercuribenzenesulfonic acid in the presence of sucrose</title><title>Plant Physiol.; (United States)</title><addtitle>Plant Physiol</addtitle><description>Broadbean (Vicia faba L.) leaf discs have been incubated with the slowly permeant thiol reagent [203Hg]-para-chloromercuribenzenesulfonic acid (PCMBS) in the presence or in the absence of sucrose, and the release of PCMBS-labeled proteins has been monitored in media containing various concentrations of urea, ethyleneglycol-bis-(β-aminoethyl ether)-N, N, N′, N′-tetraacetic acid (EGTA), sodium cholate, sodium dodecyl sulfate, Triton X-100, octylglucoside or (3-[3-cholamidopropyl)-dimethylammonio] 1-propane-sulfonate) (CHAPS). The proteins differentially labeled by PCMBS in the presence of sucrose which, on the basis of previous results, are assumed to include the sucrose carrier, were preferentially solubilized by 1% CHAPS, 1% octylglucoside, or 1% Triton X-100. Other PCMBS-labeled proteins ('background' proteins) could be partially removed by EGTA, urea, or 0.1% cholate. Sequential treatment by 10 mM EGTA and 1% CHAPS was found to give a fraction highly enriched in the differentially labeled proteins. Analysis of the specific activity of microsomal pellets suggests that the results obtained with leaf discs give a good account of what is occurring at the plasma membrane level. These data, which suggest that the proteins differentially labeled by PCMBS in the presence of sucrose are intrinsic membrane proteins, can be used to solubilize these proteins from microsomal fractions.</description><subject>550201 - Biochemistry- Tracer Techniques</subject><subject>Absorption. Translocation of ions and substances. Permeability</subject><subject>ADDITIVES</subject><subject>Agronomy. Soil science and plant productions</subject><subject>BACTERIA</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>BETA DECAY RADIOISOTOPES</subject><subject>BETA-MINUS DECAY RADIOISOTOPES</subject><subject>Biological and medical sciences</subject><subject>CARBOHYDRATES</subject><subject>CELL CONSTITUENTS</subject><subject>CELL MEMBRANES</subject><subject>Cell physiology</subject><subject>DAYS LIVING RADIOISOTOPES</subject><subject>DETERGENTS</subject><subject>DIFFERENTIAL SOLUBILIZATION</subject><subject>DISACCHARIDES</subject><subject>Economic plant physiology</subject><subject>EMULSIFIERS</subject><subject>ETIQUETADO</subject><subject>EVEN-ODD NUCLEI</subject><subject>FEUILLE</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>HEAVY NUCLEI</subject><subject>HOJAS</subject><subject>ISOTOPE APPLICATIONS</subject><subject>ISOTOPES</subject><subject>LABELLING</subject><subject>LEAVES</subject><subject>LEGUMINOSAE</subject><subject>MARQUAGE</subject><subject>Membrane and intracellular transports</subject><subject>Membrane proteins</subject><subject>MEMBRANE TRANSPORT</subject><subject>MEMBRANES</subject><subject>Membranes and Bioenergetics</subject><subject>MERCURY 203</subject><subject>MERCURY ISOTOPES</subject><subject>MICROORGANISMS</subject><subject>Molecular and cellular biology</subject><subject>NUCLEI</subject><subject>Nutrition. Photosynthesis. Respiration. Metabolism</subject><subject>OLIGOSACCHARIDES</subject><subject>ORGANIC ACIDS</subject><subject>ORGANIC COMPOUNDS</subject><subject>ORGANIC SULFUR COMPOUNDS</subject><subject>PLANTS</subject><subject>PROTEINAS</subject><subject>PROTEINE</subject><subject>PROTEINS</subject><subject>Radioactive decay</subject><subject>RADIOISOTOPES</subject><subject>Reagents</subject><subject>RESPONSE MODIFYING FACTORS</subject><subject>RHIZOBIUM</subject><subject>SACCHARIDES</subject><subject>SACCHAROSE</subject><subject>Sodium</subject><subject>SOLUBILIDAD</subject><subject>SOLUBILITE</subject><subject>SOLUBILITY</subject><subject>Solubilization</subject><subject>SUCROSA</subject><subject>SUCROSE</subject><subject>Sugars</subject><subject>SULFONIC ACIDS</subject><subject>SURFACE ACTIVE AGENTS</subject><subject>SURFACTANT</subject><subject>SURFACTANTES</subject><subject>SURFACTANTS</subject><subject>THIOL</subject><subject>THIOLS</subject><subject>TIOLES</subject><subject>TRACER TECHNIQUES</subject><subject>VICIA</subject><subject>VICIA FABA</subject><subject>WETTING AGENTS</subject><issn>0032-0889</issn><issn>1532-2548</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><recordid>eNpVkU-LFDEQxRtR3HH15FWkEcGDzJikkk76siCL_2DBw7rnkE5X72RJJ23SvTD7AfzcZphhVk8VqF9eVb1XVa8p2VBK-Kdp2ijYwEZw-qRaUQFszQRXT6sVIeVNlGrPqhc53xFCKFD-vDqjTdMIpsSq-nONHu3s7rHO0S-d8-7BzC6GOg71iGOXTMB6SnFGF3Ldu2HAhGF2xvtd7U1Xvvd1t6untd36mOKIyS7JdRgeMGBe_BCDs7Wxrq9dqOftXg0zBov7EXmxKWZ8WT0bjM_46ljPq5uvX35dfl9f_fz24_Lz1dqCUHQtB9HznjFLYQCBnZVcAQjWGSaBCxygBcPAGKlKFcICZYqjlEJw1nUUzquLg-60dCP2tlySjNdTcqNJOx2N0_93gtvq23ivKRENJ3uBdweBmGens3Uz2q2NIRQTdcNlK0hToA_HKSn-XjDPenTZovfFy7hkLQF4S1kLhfx4IPcu5ITDaRVK9D5dPU1agQZd0i3023-3f2SPcRbg_REw2Ro_lPCsyydOcgAm24K9OWB3eY7p1OZMEt7AY3swUZvbVBRurlXxsGFUKfgLF-DCIw</recordid><startdate>19870301</startdate><enddate>19870301</enddate><creator>M'Batchi, B</creator><creator>Pichelin, D</creator><creator>Delrot, S</creator><general>American Society of Plant Physiologists</general><scope>FBQ</scope><scope>IQODW</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>OTOTI</scope><scope>5PM</scope></search><sort><creationdate>19870301</creationdate><title>Selective solubilization of membrane proteins differentially labeled by p-chloromercuribenzenesulfonic acid in the presence of sucrose</title><author>M'Batchi, B ; Pichelin, D ; Delrot, S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3581-7f5d4d22c13f35ebc7483352ba27345ef393a23aa783a255c31284e775542bb13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>550201 - Biochemistry- Tracer Techniques</topic><topic>Absorption. Translocation of ions and substances. Permeability</topic><topic>ADDITIVES</topic><topic>Agronomy. Soil science and plant productions</topic><topic>BACTERIA</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>BETA DECAY RADIOISOTOPES</topic><topic>BETA-MINUS DECAY RADIOISOTOPES</topic><topic>Biological and medical sciences</topic><topic>CARBOHYDRATES</topic><topic>CELL CONSTITUENTS</topic><topic>CELL MEMBRANES</topic><topic>Cell physiology</topic><topic>DAYS LIVING RADIOISOTOPES</topic><topic>DETERGENTS</topic><topic>DIFFERENTIAL SOLUBILIZATION</topic><topic>DISACCHARIDES</topic><topic>Economic plant physiology</topic><topic>EMULSIFIERS</topic><topic>ETIQUETADO</topic><topic>EVEN-ODD NUCLEI</topic><topic>FEUILLE</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>HEAVY NUCLEI</topic><topic>HOJAS</topic><topic>ISOTOPE APPLICATIONS</topic><topic>ISOTOPES</topic><topic>LABELLING</topic><topic>LEAVES</topic><topic>LEGUMINOSAE</topic><topic>MARQUAGE</topic><topic>Membrane and intracellular transports</topic><topic>Membrane proteins</topic><topic>MEMBRANE TRANSPORT</topic><topic>MEMBRANES</topic><topic>Membranes and Bioenergetics</topic><topic>MERCURY 203</topic><topic>MERCURY ISOTOPES</topic><topic>MICROORGANISMS</topic><topic>Molecular and cellular biology</topic><topic>NUCLEI</topic><topic>Nutrition. Photosynthesis. Respiration. Metabolism</topic><topic>OLIGOSACCHARIDES</topic><topic>ORGANIC ACIDS</topic><topic>ORGANIC COMPOUNDS</topic><topic>ORGANIC SULFUR COMPOUNDS</topic><topic>PLANTS</topic><topic>PROTEINAS</topic><topic>PROTEINE</topic><topic>PROTEINS</topic><topic>Radioactive decay</topic><topic>RADIOISOTOPES</topic><topic>Reagents</topic><topic>RESPONSE MODIFYING FACTORS</topic><topic>RHIZOBIUM</topic><topic>SACCHARIDES</topic><topic>SACCHAROSE</topic><topic>Sodium</topic><topic>SOLUBILIDAD</topic><topic>SOLUBILITE</topic><topic>SOLUBILITY</topic><topic>Solubilization</topic><topic>SUCROSA</topic><topic>SUCROSE</topic><topic>Sugars</topic><topic>SULFONIC ACIDS</topic><topic>SURFACE ACTIVE AGENTS</topic><topic>SURFACTANT</topic><topic>SURFACTANTES</topic><topic>SURFACTANTS</topic><topic>THIOL</topic><topic>THIOLS</topic><topic>TIOLES</topic><topic>TRACER TECHNIQUES</topic><topic>VICIA</topic><topic>VICIA FABA</topic><topic>WETTING AGENTS</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>M'Batchi, B</creatorcontrib><creatorcontrib>Pichelin, D</creatorcontrib><creatorcontrib>Delrot, S</creatorcontrib><creatorcontrib>Centre National de la Recherche Scientifique (UA 574), Poitiers, France</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Plant Physiol.; (United States)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>M'Batchi, B</au><au>Pichelin, D</au><au>Delrot, S</au><aucorp>Centre National de la Recherche Scientifique (UA 574), Poitiers, France</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Selective solubilization of membrane proteins differentially labeled by p-chloromercuribenzenesulfonic acid in the presence of sucrose</atitle><jtitle>Plant Physiol.; (United States)</jtitle><addtitle>Plant Physiol</addtitle><date>1987-03-01</date><risdate>1987</risdate><volume>83</volume><issue>3</issue><spage>541</spage><epage>545</epage><pages>541-545</pages><issn>0032-0889</issn><eissn>1532-2548</eissn><coden>PPHYA5</coden><abstract>Broadbean (Vicia faba L.) leaf discs have been incubated with the slowly permeant thiol reagent [203Hg]-para-chloromercuribenzenesulfonic acid (PCMBS) in the presence or in the absence of sucrose, and the release of PCMBS-labeled proteins has been monitored in media containing various concentrations of urea, ethyleneglycol-bis-(β-aminoethyl ether)-N, N, N′, N′-tetraacetic acid (EGTA), sodium cholate, sodium dodecyl sulfate, Triton X-100, octylglucoside or (3-[3-cholamidopropyl)-dimethylammonio] 1-propane-sulfonate) (CHAPS). The proteins differentially labeled by PCMBS in the presence of sucrose which, on the basis of previous results, are assumed to include the sucrose carrier, were preferentially solubilized by 1% CHAPS, 1% octylglucoside, or 1% Triton X-100. Other PCMBS-labeled proteins ('background' proteins) could be partially removed by EGTA, urea, or 0.1% cholate. Sequential treatment by 10 mM EGTA and 1% CHAPS was found to give a fraction highly enriched in the differentially labeled proteins. Analysis of the specific activity of microsomal pellets suggests that the results obtained with leaf discs give a good account of what is occurring at the plasma membrane level. These data, which suggest that the proteins differentially labeled by PCMBS in the presence of sucrose are intrinsic membrane proteins, can be used to solubilize these proteins from microsomal fractions.</abstract><cop>Rockville, MD</cop><pub>American Society of Plant Physiologists</pub><pmid>16665285</pmid><doi>10.1104/pp.83.3.541</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0032-0889 |
| ispartof | Plant Physiol.; (United States), 1987-03, Vol.83 (3), p.541-545 |
| issn | 0032-0889 1532-2548 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1056401 |
| source | JSTOR Archive Collection A-Z Listing; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | 550201 - Biochemistry- Tracer Techniques Absorption. Translocation of ions and substances. Permeability ADDITIVES Agronomy. Soil science and plant productions BACTERIA BASIC BIOLOGICAL SCIENCES BETA DECAY RADIOISOTOPES BETA-MINUS DECAY RADIOISOTOPES Biological and medical sciences CARBOHYDRATES CELL CONSTITUENTS CELL MEMBRANES Cell physiology DAYS LIVING RADIOISOTOPES DETERGENTS DIFFERENTIAL SOLUBILIZATION DISACCHARIDES Economic plant physiology EMULSIFIERS ETIQUETADO EVEN-ODD NUCLEI FEUILLE Fundamental and applied biological sciences. Psychology Gels HEAVY NUCLEI HOJAS ISOTOPE APPLICATIONS ISOTOPES LABELLING LEAVES LEGUMINOSAE MARQUAGE Membrane and intracellular transports Membrane proteins MEMBRANE TRANSPORT MEMBRANES Membranes and Bioenergetics MERCURY 203 MERCURY ISOTOPES MICROORGANISMS Molecular and cellular biology NUCLEI Nutrition. Photosynthesis. Respiration. Metabolism OLIGOSACCHARIDES ORGANIC ACIDS ORGANIC COMPOUNDS ORGANIC SULFUR COMPOUNDS PLANTS PROTEINAS PROTEINE PROTEINS Radioactive decay RADIOISOTOPES Reagents RESPONSE MODIFYING FACTORS RHIZOBIUM SACCHARIDES SACCHAROSE Sodium SOLUBILIDAD SOLUBILITE SOLUBILITY Solubilization SUCROSA SUCROSE Sugars SULFONIC ACIDS SURFACE ACTIVE AGENTS SURFACTANT SURFACTANTES SURFACTANTS THIOL THIOLS TIOLES TRACER TECHNIQUES VICIA VICIA FABA WETTING AGENTS |
| title | Selective solubilization of membrane proteins differentially labeled by p-chloromercuribenzenesulfonic acid in the presence of sucrose |
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