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Our method combines two established methods: (i) the chemical modification of surface lysine residues of native proteins by <jats:italic>N<\/jats:italic>\u2010hydroxysuccinimidobiotin (NHS\u2010biotin) and (ii) the stable isotope labeling of amino acids in cell culture (SILAC) followed by tryptic digestion and mass spectrometry. By linking the chemical modification with the SILAC methodology for the first time, we obtain quantitative data on chemical modification allowing subtle changes in accessibility to be described. Five regions in the PB1 monomer showed altered reactivity to NHS\u2010biotin when compared with the [PB1\u2010PA] heterodimer. Mutational analysis of residues in two such regions\u2014at K265 and K481 of PB1, which were about three\u2010 and twofold, respectively, less accessible to biotinylation in the PB1\u2010PA heterodimer compared with the PB1 monomer, demonstrated that both K265 and K481 were crucial for polymerase function. This novel assay of quantitative profiling of biotinylation patterns (Q\u2010POP assay) highlights likely conformational changes at important functional sites, as observed here for PB1, and may provide information on protein\u2013protein interaction interfaces. The Q\u2010POP assay should be a generally applicable approach and may detect novel functional sites suitable for targeting by drugs.<\/jats:p>","DOI":"10.1002\/pro.159","type":"journal-article","created":{"date-parts":[[2009,5,13]],"date-time":"2009-05-13T21:09:31Z","timestamp":1242248971000},"page":"1448-1458","source":"Crossref","is-referenced-by-count":13,"title":["A quantitative strategy to detect changes in accessibility of protein regions to chemical modification on heterodimerization"],"prefix":"10.1002","volume":"18","author":[{"given":"Mathias","family":"Dreger","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo Wah","family":"Leung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George 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