{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T00:48:51Z","timestamp":1760575731257,"version":"build-2065373602"},"reference-count":22,"publisher":"Wiley","issue":"18","license":[{"start":{"date-parts":[[2014,7,24]],"date-time":"2014-07-24T00:00:00Z","timestamp":1406160000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["RR016464 and GM103440"],"award-info":[{"award-number":["RR016464 and GM103440"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["RR024210 and GM103554"],"award-info":[{"award-number":["RR024210 and GM103554"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[2014,9]]},"abstract":"<jats:p>Currently, there are few methods to detect differences in posttranslational modifications (PTMs) in a specific manner from complex mixtures. Thus, we developed an approach that combines the sensitivity and specificity of click chemistry with the resolution capabilities of 2D\u2010DIGE. In \u201cClick\u2010DIGE\u201d, posttranslationally modified proteins are metabolically labeled with azido\u2010substrate analogs, then size\u2010 and charge\u2010matched alkyne\u2010Cy3 or alkyne\u2010Cy5 dyes are covalently attached to the azide of the PTM by click chemistry. The fluorescently\u2010tagged protein samples are then multiplexed for 2DE analysis. Whereas standard DIGE labels all proteins, Click\u2010DIGE focuses the analysis of protein differences to a targeted subset of posttranslationally modified proteins within a complex sample (i.e. specific labeling and analysis of azido glycoproteins within a cell lysate). Our data indicate that (i) Click\u2010DIGE specifically labels azido proteins, (ii) the resulting Cy\u2010protein conjugates are spectrally distinct, and (iii) the conjugates are size\u2010 and charge\u2010matched at the level of 2DE. We demonstrate the utility of this approach by detecting multiple differentially expressed glycoproteins between a mutant cell line defective in UDP\u2010galactose transport and the parental cell line. We anticipate that the diversity of azido substrates already available will enable Click\u2010DIGE to be compatible with analysis of a wide range of PTMs.<\/jats:p>","DOI":"10.1002\/elps.201400241","type":"journal-article","created":{"date-parts":[[2014,6,16]],"date-time":"2014-06-16T18:33:33Z","timestamp":1402943613000},"page":"2621-2625","source":"Crossref","is-referenced-by-count":4,"title":["Size\u2010matched alkyne\u2010conjugated cyanine fluorophores to identify differences in protein glycosylation"],"prefix":"10.1002","volume":"35","author":[{"given":"Amanda R.","family":"Burnham\u2010Marusich","sequence":"first","affiliation":[{"name":"Biology Department University of Nevada  Reno NV USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna M.","family":"Plechaty","sequence":"additional","affiliation":[{"name":"Biology Department University of Nevada  Reno NV USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia M.","family":"Berninsone","sequence":"additional","affiliation":[{"name":"Biology Department University of Nevada  Reno NV USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2014,7,24]]},"reference":[{"key":"e_1_2_4_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.1150181133"},{"key":"e_1_2_4_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200390006"},{"key":"e_1_2_4_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/1615-9861(200103)1:3<377::AID-PROT377>3.0.CO;2-6"},{"key":"e_1_2_4_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200300439"},{"key":"e_1_2_4_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2013.08.048"},{"volume-title":"Chemistry","year":"2008","author":"Fernando M. 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