Differential recovery of membrane proteins after extraction by aqueous methanol and trifluoroethanol
Cell membrane proteome analysis is limited by inherent membrane hydrophobicity. Conventional membrane protein extraction techniques use detergents, chaotropes and organic acids that require sample clean‐up or pH adjustment, and are associated with significant sample loss. We extracted membrane prote...
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| Veröffentlicht in: | Proteomics (Weinheim) 2007-05, Vol.7 (10), p.1654-1663 |
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| creator | Zhang, Huoming Lin, Qingsong Ponnusamy, Sukumar Kothandaraman, Narasimhan Lim, Teck Kwang Zhao, Changqing Kit, Hon Sook Arijit, Biswas Rauff, Mary Hew, Choy-Leong Chung, Maxey Ching Ming Joshi, Shashikant B. Choolani, Mahesh |
| description | Cell membrane proteome analysis is limited by inherent membrane hydrophobicity. Conventional membrane protein extraction techniques use detergents, chaotropes and organic acids that require sample clean‐up or pH adjustment, and are associated with significant sample loss. We extracted membrane proteins from red blood cells (RBCs) using methanol (MeOH), trifluoroethanol (TFE) and urea, and identified membrane proteins using 2‐D LC coupled with MALDI‐TOF/TOF‐MS. We show that organic solvents MeOH‐ and TFE‐based methods have membrane protein analysis efficiencies comparable to urea, and are complementary for the recovery of both hydrophilic and hydrophobic peptides. The mean grand average of hydropathicity (GRAVY) value of identified peptides from the TFE‐based method (−0.107) was significantly higher than that of the MeOH‐based method (−0.465) (p |
| doi_str_mv | 10.1002/pmic.200600579 |
| format | Article |
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Conventional membrane protein extraction techniques use detergents, chaotropes and organic acids that require sample clean‐up or pH adjustment, and are associated with significant sample loss. We extracted membrane proteins from red blood cells (RBCs) using methanol (MeOH), trifluoroethanol (TFE) and urea, and identified membrane proteins using 2‐D LC coupled with MALDI‐TOF/TOF‐MS. We show that organic solvents MeOH‐ and TFE‐based methods have membrane protein analysis efficiencies comparable to urea, and are complementary for the recovery of both hydrophilic and hydrophobic peptides. The mean grand average of hydropathicity (GRAVY) value of identified peptides from the TFE‐based method (−0.107) was significantly higher than that of the MeOH‐based method (−0.465) (p<0.001). Sequential and adjunctive use of the organic solvents MeOH and TFE increases the number of proteins identified, and the confidence of their identification. We show that this strategy is effective for shotgun membrane proteome analysis.</description><identifier>ISSN: 1615-9853</identifier><identifier>EISSN: 1615-9861</identifier><identifier>DOI: 10.1002/pmic.200600579</identifier><identifier>PMID: 17436264</identifier><language>eng</language><publisher>Weinheim: WILEY-VCH Verlag</publisher><subject>2-D LC-MALDI-TOF/TOF-MS ; Adult ; Cell Membrane - chemistry ; Chromatography, Liquid ; Electrophoresis, Gel, Two-Dimensional ; Erythrocytes - chemistry ; Human erythrocyte membrane protein ; Humans ; Mass Spectrometry ; Membrane Proteins - chemistry ; Membrane Proteins - isolation & purification ; Methanol ; Methanol - chemistry ; Molecular Sequence Data ; Solvents - chemistry ; Trifluoroethanol ; Trifluoroethanol - chemistry</subject><ispartof>Proteomics (Weinheim), 2007-05, Vol.7 (10), p.1654-1663</ispartof><rights>Copyright © 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4479-839b254b323379dac5f7681584c8e6dd73213c72e56ca822b03d2bc6c1e70b13</citedby><cites>FETCH-LOGICAL-c4479-839b254b323379dac5f7681584c8e6dd73213c72e56ca822b03d2bc6c1e70b13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fpmic.200600579$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fpmic.200600579$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17436264$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Huoming</creatorcontrib><creatorcontrib>Lin, Qingsong</creatorcontrib><creatorcontrib>Ponnusamy, Sukumar</creatorcontrib><creatorcontrib>Kothandaraman, Narasimhan</creatorcontrib><creatorcontrib>Lim, Teck Kwang</creatorcontrib><creatorcontrib>Zhao, Changqing</creatorcontrib><creatorcontrib>Kit, Hon Sook</creatorcontrib><creatorcontrib>Arijit, Biswas</creatorcontrib><creatorcontrib>Rauff, Mary</creatorcontrib><creatorcontrib>Hew, Choy-Leong</creatorcontrib><creatorcontrib>Chung, Maxey Ching Ming</creatorcontrib><creatorcontrib>Joshi, Shashikant B.</creatorcontrib><creatorcontrib>Choolani, Mahesh</creatorcontrib><title>Differential recovery of membrane proteins after extraction by aqueous methanol and trifluoroethanol</title><title>Proteomics (Weinheim)</title><addtitle>Proteomics</addtitle><description>Cell membrane proteome analysis is limited by inherent membrane hydrophobicity. Conventional membrane protein extraction techniques use detergents, chaotropes and organic acids that require sample clean‐up or pH adjustment, and are associated with significant sample loss. We extracted membrane proteins from red blood cells (RBCs) using methanol (MeOH), trifluoroethanol (TFE) and urea, and identified membrane proteins using 2‐D LC coupled with MALDI‐TOF/TOF‐MS. We show that organic solvents MeOH‐ and TFE‐based methods have membrane protein analysis efficiencies comparable to urea, and are complementary for the recovery of both hydrophilic and hydrophobic peptides. The mean grand average of hydropathicity (GRAVY) value of identified peptides from the TFE‐based method (−0.107) was significantly higher than that of the MeOH‐based method (−0.465) (p<0.001). Sequential and adjunctive use of the organic solvents MeOH and TFE increases the number of proteins identified, and the confidence of their identification. We show that this strategy is effective for shotgun membrane proteome analysis.</description><subject>2-D LC-MALDI-TOF/TOF-MS</subject><subject>Adult</subject><subject>Cell Membrane - chemistry</subject><subject>Chromatography, Liquid</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Erythrocytes - chemistry</subject><subject>Human erythrocyte membrane protein</subject><subject>Humans</subject><subject>Mass Spectrometry</subject><subject>Membrane Proteins - chemistry</subject><subject>Membrane Proteins - isolation & purification</subject><subject>Methanol</subject><subject>Methanol - chemistry</subject><subject>Molecular Sequence Data</subject><subject>Solvents - chemistry</subject><subject>Trifluoroethanol</subject><subject>Trifluoroethanol - chemistry</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkElPwzAQhS0EYr9yRD5xS_ES28kRlVUqixAS3CzHmQhDEhfbhfbfE9SqcOM0o9H3nt48hI4oGVFC2Om0c3bECJGECFVuoF0qqcjKQtLN9S74DtqL8Y0QqopSbaMdqnIumcx3UX3umgYC9MmZFgew_hPCAvsGd9BVwfSAp8EncH3EpkkQMMxTMDY53-Nqgc3HDPwsDnR6Nb1vselrnIJr2pkPfnU8QFuNaSMcruY-erq8eBpfZ5P7q5vx2SSzea7KrOBlxURecca5KmtjRaNkQUWR2wJkXSvOKLeKgZDWFIxVhNesstJSUKSifB-dLG2HxEOsmHTnooW2Hb4YMmpFBBGMsgEcLUEbfIwBGj0NrjNhoSnRP7Xqn1r1utZBcLxynlUd1L_4qscBKJfAl2th8Y-dfri9Gf81z5ZaFxPM11oT3rVUXAn9fHelxYsoHyl51rf8G2PMlUU</recordid><startdate>20070501</startdate><enddate>20070501</enddate><creator>Zhang, Huoming</creator><creator>Lin, Qingsong</creator><creator>Ponnusamy, Sukumar</creator><creator>Kothandaraman, Narasimhan</creator><creator>Lim, Teck Kwang</creator><creator>Zhao, Changqing</creator><creator>Kit, Hon Sook</creator><creator>Arijit, Biswas</creator><creator>Rauff, Mary</creator><creator>Hew, Choy-Leong</creator><creator>Chung, Maxey Ching Ming</creator><creator>Joshi, Shashikant B.</creator><creator>Choolani, Mahesh</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><scope>BSCLL</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>20070501</creationdate><title>Differential recovery of membrane proteins after extraction by aqueous methanol and trifluoroethanol</title><author>Zhang, Huoming ; Lin, Qingsong ; Ponnusamy, Sukumar ; Kothandaraman, Narasimhan ; Lim, Teck Kwang ; Zhao, Changqing ; Kit, Hon Sook ; Arijit, Biswas ; Rauff, Mary ; Hew, Choy-Leong ; Chung, Maxey Ching Ming ; Joshi, Shashikant B. ; Choolani, Mahesh</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4479-839b254b323379dac5f7681584c8e6dd73213c72e56ca822b03d2bc6c1e70b13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>2-D LC-MALDI-TOF/TOF-MS</topic><topic>Adult</topic><topic>Cell Membrane - chemistry</topic><topic>Chromatography, Liquid</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Erythrocytes - chemistry</topic><topic>Human erythrocyte membrane protein</topic><topic>Humans</topic><topic>Mass Spectrometry</topic><topic>Membrane Proteins - chemistry</topic><topic>Membrane Proteins - isolation & purification</topic><topic>Methanol</topic><topic>Methanol - chemistry</topic><topic>Molecular Sequence Data</topic><topic>Solvents - chemistry</topic><topic>Trifluoroethanol</topic><topic>Trifluoroethanol - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Huoming</creatorcontrib><creatorcontrib>Lin, Qingsong</creatorcontrib><creatorcontrib>Ponnusamy, Sukumar</creatorcontrib><creatorcontrib>Kothandaraman, Narasimhan</creatorcontrib><creatorcontrib>Lim, Teck Kwang</creatorcontrib><creatorcontrib>Zhao, Changqing</creatorcontrib><creatorcontrib>Kit, Hon Sook</creatorcontrib><creatorcontrib>Arijit, Biswas</creatorcontrib><creatorcontrib>Rauff, Mary</creatorcontrib><creatorcontrib>Hew, Choy-Leong</creatorcontrib><creatorcontrib>Chung, Maxey Ching Ming</creatorcontrib><creatorcontrib>Joshi, Shashikant B.</creatorcontrib><creatorcontrib>Choolani, Mahesh</creatorcontrib><collection>Istex</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>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Huoming</au><au>Lin, Qingsong</au><au>Ponnusamy, Sukumar</au><au>Kothandaraman, Narasimhan</au><au>Lim, Teck Kwang</au><au>Zhao, Changqing</au><au>Kit, Hon Sook</au><au>Arijit, Biswas</au><au>Rauff, Mary</au><au>Hew, Choy-Leong</au><au>Chung, Maxey Ching Ming</au><au>Joshi, Shashikant B.</au><au>Choolani, Mahesh</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Differential recovery of membrane proteins after extraction by aqueous methanol and trifluoroethanol</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2007-05-01</date><risdate>2007</risdate><volume>7</volume><issue>10</issue><spage>1654</spage><epage>1663</epage><pages>1654-1663</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>Cell membrane proteome analysis is limited by inherent membrane hydrophobicity. Conventional membrane protein extraction techniques use detergents, chaotropes and organic acids that require sample clean‐up or pH adjustment, and are associated with significant sample loss. We extracted membrane proteins from red blood cells (RBCs) using methanol (MeOH), trifluoroethanol (TFE) and urea, and identified membrane proteins using 2‐D LC coupled with MALDI‐TOF/TOF‐MS. We show that organic solvents MeOH‐ and TFE‐based methods have membrane protein analysis efficiencies comparable to urea, and are complementary for the recovery of both hydrophilic and hydrophobic peptides. The mean grand average of hydropathicity (GRAVY) value of identified peptides from the TFE‐based method (−0.107) was significantly higher than that of the MeOH‐based method (−0.465) (p<0.001). Sequential and adjunctive use of the organic solvents MeOH and TFE increases the number of proteins identified, and the confidence of their identification. We show that this strategy is effective for shotgun membrane proteome analysis.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>17436264</pmid><doi>10.1002/pmic.200600579</doi><tpages>10</tpages></addata></record> |
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| subjects | 2-D LC-MALDI-TOF/TOF-MS Adult Cell Membrane - chemistry Chromatography, Liquid Electrophoresis, Gel, Two-Dimensional Erythrocytes - chemistry Human erythrocyte membrane protein Humans Mass Spectrometry Membrane Proteins - chemistry Membrane Proteins - isolation & purification Methanol Methanol - chemistry Molecular Sequence Data Solvents - chemistry Trifluoroethanol Trifluoroethanol - chemistry |
| title | Differential recovery of membrane proteins after extraction by aqueous methanol and trifluoroethanol |
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