{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T12:24:37Z","timestamp":1788870277948,"version":"build-2803163510"},"reference-count":33,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2011,8,18]],"date-time":"2011-08-18T00:00:00Z","timestamp":1313625600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Carl Tryggers Foundation for Scientific Research","award":["CTS09:96"],"award-info":[{"award-number":["CTS09:96"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2011,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The lysine\u2010specific crosslinker 3,3\u2032\u2010dithiobis(sulfosuccinimidylpropionate) (DTSSP) is commonly used in the structural characterization of proteins by chemical crosslinking and mass spectrometry and we here describe an efficient two\u2010step LC\u2010MALDI\u2010TOF\/TOF procedure to detect crosslinked peptides. First MS data are acquired, and the properties of isotope\u2010labeled DTSSP are used in data analysis to identify candidate crosslinks. MSMS data are then acquired for a restricted number of precursor ions per spot for final crosslink identification. We show that the thiol\u2010catalyzed exchange between crosslinked peptides, which is due to the disulfide bond in DTSSP and known to possibly obscure data, can be precisely quantified using isotope\u2010labeled DTSSP. Crosslinked peptides are recognized as 8 Da doublet peaks and a new isotopic peak with twice the intensity appears in the middle of the doublet as a consequence of the thiol\u2010exchange. False\u2010positive crosslinks, formed exclusively by thiol\u2010exchange, yield a 1:2:1 isotope pattern, whereas true crosslinks, formed by two lysine residues within crosslinkable distance in the native protein structure, yield a 1:0:1 isotope pattern. Peaks with a 1:\n                    <jats:italic>X<\/jats:italic>\n                    :1 isotope pattern, where 0 &lt;\n                    <jats:italic>X<\/jats:italic>\n                    &lt; 2, can be trusted as true crosslinks, with a defined proportion of the signal [2\n                    <jats:italic>X<\/jats:italic>\n                    \/(2 +\n                    <jats:italic>X<\/jats:italic>\n                    )] being noise from the thiol\u2010exchange. The thiol\u2010exchange was correlated with the protein cysteine content and was minimized by shortening the trypsin incubation time, and for two molecular chaperone proteins with known structure all crosslinks fitted well to the structure data. The thiol\u2010exchange can thus be controlled and isotope\u2010labeled DTSSP safely used to detect true crosslinks between lysine residues in proteins.\n                  <\/jats:p>","DOI":"10.1002\/pro.699","type":"journal-article","created":{"date-parts":[[2011,7,20]],"date-time":"2011-07-20T16:01:46Z","timestamp":1311177706000},"page":"1682-1691","source":"Crossref","is-referenced-by-count":6,"title":["Thiol\u2010exchange in DTSSP crosslinked peptides is proportional to cysteine content and precisely controlled in crosslink detection by two\u2010step LC\u2010MALDI MSMS"],"prefix":"10.1002","volume":"20","author":[{"given":"Wietske","family":"Lambert","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher A. G.","family":"S\u00f6derberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gudrun","family":"Rutsdottir","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wilbert C.","family":"Boelens","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cecilia","family":"Emanuelsson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2011,8,18]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.R000001-MCP201"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac1000724"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/mas.20082"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1039\/b801810c"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.1144"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac900853k"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2009.401"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac001379a"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac0488762"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M800232-MCP200"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac101241t"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac101586z"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac901637v"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac0257492"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac702277q"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.9.8.1503"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.0212602"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.04910004"},{"key":"e_1_2_6_20_2","first-page":"857","article-title":"Probing alpha\u2010crystallin structure using chemical cross\u2010linkers and mass spectrometry","volume":"10","author":"Peterson JJ","year":"2004","journal-title":"Mol Vis"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.072831607"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/mas.10025"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.560"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01751808"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.molbev.a026406"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.07.069"},{"key":"e_1_2_6_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.380"},{"key":"e_1_2_6_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb.1891"},{"key":"e_1_2_6_29_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0902651106"},{"key":"e_1_2_6_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0141-8130(98)00017-8"},{"key":"e_1_2_6_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-4838(00)00243-0"},{"key":"e_1_2_6_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_2_6_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(90)82008-P"},{"key":"e_1_2_6_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(01)01954-5"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fpro.699","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fpro.699","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.699","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,9]],"date-time":"2023-10-09T11:19:09Z","timestamp":1696850349000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.699"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,8,18]]},"references-count":33,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2011,10]]}},"alternative-id":["10.1002\/pro.699"],"URL":"https:\/\/doi.org\/10.1002\/pro.699","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,8,18]]}}}