{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T12:50:29Z","timestamp":1761483029810,"version":"build-2065373602"},"reference-count":36,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2009,2,9]],"date-time":"2009-02-09T00:00:00Z","timestamp":1234137600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2009,3,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The rapid and specific detection of therapeutically important ligands in complex mixtures, that may bind to membrane proteins, remains challenging for many research laboratories and pharmaceutical industries. Through its use in the development of screening assays, mass spectrometry (MS) is currently experiencing a period of tremendous expansion. In the study presented here, we took advantage of the remarkable stability properties of a bacterial membrane protein, the KcsA K\n                    <jats:sup>+<\/jats:sup>\n                    channel, produced in\n                    <jats:italic>E<\/jats:italic>\n                    .\n                    <jats:italic>coli<\/jats:italic>\n                    and purified as a tetrameric protein in the presence of a detergent. This membrane protein can subserve as a molecular template to display the pore\u2010forming region of human K\n                    <jats:sup>+<\/jats:sup>\n                    channels, which are considered as targets in the search for inhibitory ligands. The engineered chimeric proteins were linked to metal\u2010bound magnetic beads, for the screening of complex peptide mixtures, such as that of scorpion venoms. The affinity\u2010captured scorpion toxins were eluted prior to matrix\u2010assisted laser desorption\/ionization time\u2010of\u2010flight mass spectrometry (MALDI\u2010TOFMS), and to nano\u2010electrospray ionization tandem mass QqTOF mass spectrometry (MS\/MS) analysis. The\n                    <jats:italic>de novo<\/jats:italic>\n                    sequence of the toxins was deduced by combining the MS\/MS fragmentation of the reduced form (up to the 33 first residues) and the trypsin digest peptides of the native toxins. This affinity\u2010capture screening assay led to the isolation and characterization of potent and specific ligands of the human K\n                    <jats:sup>+<\/jats:sup>\n                    channel, Kv1.3. The affinity\u2010capture procedure is fast and reproducible. When linked to magnetic beads, the chimeric membrane protein can be re\u2010used several times without losing any of its selectivity or specificity. This assay also benefits from the fact that it requires minimal amounts of animal venoms or complex mixtures, which can be expensive or difficult to procure. Copyright \u00a9 2009 John Wiley &amp; Sons, Ltd.\n                  <\/jats:p>","DOI":"10.1002\/rcm.3939","type":"journal-article","created":{"date-parts":[[2009,2,9]],"date-time":"2009-02-09T06:14:21Z","timestamp":1234160061000},"page":"745-755","source":"Crossref","is-referenced-by-count":5,"title":["Affinity capture using chimeric membrane proteins bound to magnetic beads for rapid ligand screening by matrix\u2010assisted laser desorption\/ionization time\u2010of\u2010flight mass spectrometry"],"prefix":"10.1002","volume":"23","author":[{"given":"Christian","family":"Legros","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catherine","family":"Guette","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie\u2010France","family":"Martin\u2010Eauclaire","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Max","family":"Goyffon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeanine","family":"Tortajada","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2009,2,9]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/mas.20019"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2005.08.038"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2007.08.031"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac00291a042"},{"issue":"1","key":"e_1_2_6_6_2","first-page":"193A","volume":"63","author":"Hillenkamp F","year":"1991","journal-title":"Anal. 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