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In this study, we combined chemical cross\u2010linking with mass spectrometry to determine the binding stoichiometry and map the protein\u2013protein interaction network of a human SAGA HAT subcomplex. MALDI\u2010MS equipped with high mass detection was used to follow the cross\u2010linking reaction using bis[sulfosuccinimidyl] suberate (BS3) and confirm the heterotetrameric stoichiometry of the specific stabilized subcomplex. Cross\u2010linking with isotopically labeled BS3 d0\u2010d4 followed by trypsin digestion allowed the identification of intra\u2010 and intercross\u2010linked peptides using two dedicated search engines: pLink and xQuest. The identified interlinked peptides suggest a strong network of interaction between GCN5, ADA2B and ADA3 subunits; SGF29 is interacting with GCN5 and ADA3 but not with ADA2B. These restraint data were combined to molecular modeling and a low\u2010resolution interacting model for the human SAGA HAT subcomplex could be proposed, illustrating the potential of an integrative strategy using cross\u2010linking and mass spectrometry for addressing the structural architecture of multiprotein complexes.<\/jats:p>","DOI":"10.1002\/pro.2676","type":"journal-article","created":{"date-parts":[[2015,3,9]],"date-time":"2015-03-09T09:18:57Z","timestamp":1425892737000},"page":"1232-1246","source":"Crossref","is-referenced-by-count":25,"title":["Chemical cross\u2010linking and mass spectrometry to determine the subunit interaction network in a recombinant human SAGA HAT subcomplex"],"prefix":"10.1002","volume":"24","author":[{"given":"Nha\u2010Thi","family":"Nguyen\u2010Huynh","sequence":"first","affiliation":[{"name":"Laboratoire de Spectrom\u00e9trie de Masse des Interactions et des Syst\u00e8mes (LSMIS) UMR 7140 CNRS\/Universit\u00e9 de Strasbourg \u2010 \"Chimie de la 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