{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T04:30:26Z","timestamp":1761539426723,"version":"build-2065373602"},"reference-count":7,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2009,7,6]],"date-time":"2009-07-06T00:00:00Z","timestamp":1246838400000},"content-version":"vor","delay-in-days":35,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[2009,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Knowledge on complete sequences is pivotal to identify splice variants, generate specific antibodies and predict conformation. A simple analytical approach to obtain 100% sequence coverage, however, is currently not available. Recombinant gamma\u2010aminobutyric acid A receptor subunits were from insect SF9 cells that were co\u2010transfected with rat \u03b11 and His\u2010tag \u03b23. The complex of these two subunits was run on blue\u2010native PAGE, followed by multidimensional electrophoretic steps. Spots resolved at the third electrophoretic step were in\u2010gel digested with trypsin, chymotrypsin and Asp\u2010N. In\u2010gel modification of lysines by acetylation was carried out to increase sequence coverage. Subsequently, peptides were analyzed by nano\u2010ESI\u2010LC\u2010MS\/MS using both, collision\u2010induced dissociation and electron transfer dissociation principles. When results from trypsin, chymotrypsin and Asp\u2010N digestion were combined, a single peptide [424 KKTHLRRRSSQL 435] was still not identified. In\u2010gel lysine acetylation leads to unambiguous identification of this peptide by the use of MASCOT v2.2. The use of the Modiro<jats:sup>\u2122<\/jats:sup> software along with MASCOT, however, was able to provide 100% sequence coverage even without the use of in\u2010gel lysine acetylation. It was observed that the use of trypsin, chymotrypsin and Asp\u2010N with bioinformatic handling by the MASCOT and Modiro<jats:sup>\u2122<\/jats:sup> software is sufficient to obtain complete sequencing of a highly hydrophobic membrane protein.<\/jats:p>","DOI":"10.1002\/elps.200900024","type":"journal-article","created":{"date-parts":[[2009,7,6]],"date-time":"2009-07-06T21:54:16Z","timestamp":1246917256000},"page":"2159-2167","source":"Crossref","is-referenced-by-count":7,"title":["Complete sequencing of GABA<sub>A<\/sub> receptor subunit \u03b23 by a rapid technique following in\u2010gel digestion of the protein"],"prefix":"10.1002","volume":"30","author":[{"given":"Sung Ung","family":"Kang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gert","family":"Lubec","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2009,7,6]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/S0031-6997(25)06840-1","volume":"47","author":"Sieghart W","year":"1995","journal-title":"Pharmacol. Rev."},{"key":"e_1_2_6_3_2","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S0031-6997(24)01363-2","volume":"50","author":"Barnard E. A.","year":"1998","journal-title":"Pharmacol. Rev."},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.2174\/1568026023393507"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M401354200"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr800236u"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.7"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200600189"}],"container-title":["ELECTROPHORESIS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Felps.200900024","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/elps.200900024","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T17:17:18Z","timestamp":1760548638000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/elps.200900024"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,6]]},"references-count":7,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2009,6]]}},"alternative-id":["10.1002\/elps.200900024"],"URL":"https:\/\/doi.org\/10.1002\/elps.200900024","archive":["Portico"],"relation":{},"ISSN":["0173-0835","1522-2683"],"issn-type":[{"type":"print","value":"0173-0835"},{"type":"electronic","value":"1522-2683"}],"subject":[],"published":{"date-parts":[[2009,6]]}}}