{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T23:21:44Z","timestamp":1777072904944,"version":"3.51.4"},"reference-count":25,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2013,6,18]],"date-time":"2013-06-18T00:00:00Z","timestamp":1371513600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Mass Spectrom."],"published-print":{"date-parts":[[2013,7]]},"abstract":"<jats:p>Chemical cross\u2010linking, combined with mass spectrometry, has been applied to map three\u2010dimensional protein structures and protein\u2013protein interactions. Proper choice of the cross\u2010linking agent, including its reactive groups and spacer arm length, is of great importance. However, studies to understand the details of reactivity of the chemical cross\u2010linkers with proteins are quite sparse. In this study, we investigated chemical cross\u2010linking from the aspects of the protein structures and the cross\u2010linking reagents involved, by using two structurally well\u2010known proteins, glyceraldehyde 3\u2010phosohate dehydrogenase and ribonuclease S. Chemical cross\u2010linking reactivity was compared using a series of homo\u2010 and hetero\u2010bifunctional cross\u2010linkers, including bis(sulfosuccinimidyl) suberate, dissuccinimidyl suberate, bis(succinimidyl) penta (ethylene glycol), bis(succinimidyl) nona (ethylene glycol), <jats:italic>m<\/jats:italic>\u2010maleimidobenzoyl\u2010<jats:italic>N<\/jats:italic>\u2010hydroxysulfosuccinimide ester, 2\u2010pyridyldithiol\u2010tetraoxaoctatriacontane\u2010<jats:italic>N<\/jats:italic>\u2010hydrosuccinimide and succinimidyl\u2010[(<jats:italic>N<\/jats:italic>\u2010maleimidopropionamido)\u2010tetracosaethyleneglycol]ester. The protein structure itself, especially the distances between target amino acid residues, was found to be a determining factor for the cross\u2010linking efficiency. Moreover, the reactive groups of the chemical cross\u2010linker also play an important role; a higher cross\u2010linking reaction efficiency was found for maleimides compared to 2\u2010pyrimidyldithiols. The reaction between maleimides and sulfhydryl groups is more favorable than that between <jats:italic>N<\/jats:italic>\u2010hydroxysuccinimide esters and amine groups, although cysteine residues are less abundant in proteins compared to lysine residues. Copyright \u00a9 2013 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jms.3224","type":"journal-article","created":{"date-parts":[[2013,6,27]],"date-time":"2013-06-27T05:16:13Z","timestamp":1372310173000},"page":"807-812","source":"Crossref","is-referenced-by-count":36,"title":["Understanding chemical reactivity for homo\u2010 and heterobifunctional protein cross\u2010linking agents"],"prefix":"10.1002","volume":"48","author":[{"given":"Fan","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Chemistry and Applied Biosciences ETH Z\u00fcrich  CH\u20108093 Z\u00fcrich Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simone","family":"Nielsen","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Applied Biosciences ETH Z\u00fcrich  CH\u20108093 Z\u00fcrich Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Renato","family":"Zenobi","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Applied Biosciences ETH Z\u00fcrich  CH\u20108093 Z\u00fcrich Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2013,6,18]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/mas.20082"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/jms.559"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.R000001-MCP201"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac0519108"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.200800625"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/jms.2989"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.62.070193.002411"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac1000724"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.2174\/1389203043379701"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1096-9888(199808)33:8<697::AID-JMS711>3.0.CO;2-H"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/bms.1200201209"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-9861(75)90467-1"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(76)90011-5"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/jms.1544"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-9861(66)90079-8"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja01619a073"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj0910589"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(82)90281-0"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1210074"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(72)80175-3"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj1730723"},{"key":"e_1_2_6_23_1","volume-title":"Chemistry of protein conjugation and cross\u2010linking","author":"Wong S. 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