- 第3日 5月17日（木） 10:00～10:15 A会場（オービットホール）
We developed the nucleus separation protocol by using ethylene glycol and MS-compatible surfactants. The protocol was not required the dounce homogenizer, the density gradient ultracentrifugation or/and the RIPA buffer. Nucleus were separated from AML3, HEK and Caco2 in the duplicate. Proteins were extracted with the phase transfer surfactant using sodium deoxycholate and sodium lauroyl sarcosinate. Digested peptides were analyzed by data independent acquisition mass spectrometry. In total, 1254 proteins were quantified. By using the keywords of Uniprot database, 624 nucleus proteins and 197 transcription factors were annotated. More than 25% of nuclear proteome comprises the housekeeping proteins such as histone and lamin proteins based on the MS signal. The expression level of the housekeeping nuclear proteins was identical within three cell lines. On the other hand, 10, 23 and 10 transcription factors were uniquely observed in AML3, Caco2 and HEK cells, respectively. This dataset contains some cell type specific factors.