{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T10:45:03Z","timestamp":1790246703035,"version":"4.1.0"},"reference-count":32,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2013,3,1]],"date-time":"2013-03-01T00:00:00Z","timestamp":1362096000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CP Chemical Biology"],"published-print":{"date-parts":[[2013,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Enzymatic transmethylation from the cofactor <jats:italic>S<\/jats:italic>\u2010adenosyl\u2010<jats:sc>L<\/jats:sc>\u2010methionine (SAM) to biological molecules has recently garnered increased attention because of the diversity of possible substrates and implications in normal biology and diseases. To reveal the substrates of protein methyltransferases (PMTs), the present article focuses on an alkyne\u2010containing SAM mimic, <jats:italic>Se<\/jats:italic>\u2010adenosyl\u2010<jats:sc>L<\/jats:sc>\u2010selenomethionine (ProSeAM), and a cleavable azido\u2010azo\u2010biotin probe to profile the targets of endogenous PMTs in cellular contexts. This article describes the stepwise preparation of cell lysates containing active, endogenous PMTs and subsequent target labeling with ProSeAM. The article continues with the enrichment of the ProSeAM\u2010labeled proteins with the azido\u2010azo biotin probe as a pulldown reagent and the subsequent reductive elution with sodium dithionate for proteomic analysis. The protocols provided here were formulated for ProSeAM as a profiling reagent but can be applied to other terminal\u2010alkyne\u2010containing SAM analog cofactors. <jats:italic>Curr. Protoc. Chem. Biol<\/jats:italic>. 5:67\u201088 \u00a9 2013 by John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/9780470559277.ch120241","type":"journal-article","created":{"date-parts":[[2013,3,11]],"date-time":"2013-03-11T13:51:51Z","timestamp":1363009911000},"page":"67-88","source":"Crossref","is-referenced-by-count":22,"title":["Profiling Protein Methylation with Cofactor Analog Containing Terminal Alkyne Functionality"],"prefix":"10.1002","volume":"5","author":[{"given":"Gil","family":"Blum","sequence":"first","affiliation":[{"name":"Molecular Pharmacology and Chemistry Program, Memorial Sloan\u2010Kettering Cancer Center  New York New York"},{"name":"Tri\u2010Institutional Training Program in Chemical Biology, Memorial Sloan\u2010Kettering Cancer Center  New York New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian R.","family":"Bothwell","sequence":"additional","affiliation":[{"name":"Molecular Pharmacology and Chemistry Program, Memorial Sloan\u2010Kettering Cancer Center  New York New York"},{"name":"Tri\u2010Institutional Training Program in Chemical Biology, Memorial Sloan\u2010Kettering Cancer Center  New York New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kabirul","family":"Islam","sequence":"additional","affiliation":[{"name":"Molecular Pharmacology and Chemistry Program, Memorial Sloan\u2010Kettering Cancer Center  New York New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minkui","family":"Luo","sequence":"additional","affiliation":[{"name":"Molecular Pharmacology and Chemistry Program, Memorial Sloan\u2010Kettering Cancer Center  New York New York"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,3]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature08822"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi9007726"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201000433"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780470559277.ch120240"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja304782r"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2009.07.010"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb1002as97"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M502458200"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc0602228"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2010.03.010"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.2174\/138620712800194468"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb2000567"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja2118333"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1186\/1756-8935-4-19"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb200519y"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142700.nca03es03"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201001240"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.08.017"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1039\/b811673c"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.88"},{"key":"e_1_2_12_22_1","first-page":"10.6.1","volume":"85","author":"Sasse J.","year":"2009","journal-title":"Staining proteins in gels"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb1001as76"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja1083909"},{"key":"e_1_2_12_25_1","doi-asserted-by":"crossref","first-page":"2793","DOI":"10.1016\/S0021-9258(17)40530-8","article-title":"Identification of a gamma\u2010glutamyl methyl ester in bacterial membrane protein involved in chemotaxis","volume":"252","author":"Werf P.","year":"1977","journal-title":"J. Biol. Chem."},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.599"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200501023"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.1039\/c1mb05230f"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja2006719"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201100781"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja908871t"},{"key":"e_1_2_12_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.405"},{"key":"e_1_2_12_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature10690"}],"container-title":["Current Protocols in Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2F9780470559277.ch120241","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/9780470559277.ch120241","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,24]],"date-time":"2023-09-24T11:38:42Z","timestamp":1695555522000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/9780470559277.ch120241"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,3]]},"references-count":32,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2013,3]]}},"alternative-id":["10.1002\/9780470559277.ch120241"],"URL":"https:\/\/doi.org\/10.1002\/9780470559277.ch120241","archive":["Portico"],"relation":{},"ISSN":["2160-4762","2160-4762"],"issn-type":[{"value":"2160-4762","type":"print"},{"value":"2160-4762","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,3]]}}}