日本質量分析学会 第66回質量分析総合討論会

Program

Oral Sessions

Day 3, May 17(Thu.) 10:00-10:15 Room A (OrBit Hall)

Nuclear Enrichment toward Large Scale Profiling of Transcription Factors

(1Kumamoto Univ., 2Kumamoto Univ., 3AMED-CREST)
oTakeshi Masuda1,2,3, Shingo Ito1,2,3, Sumio Ohtsuki1,2,3

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.