Dataset Information

Quantitative dissection and stoichiometry determination of the human SET1/MLL histone methyltransferase complexes

ABSTRACT: Methylation of lysine 4 at histone H3 (H3K4) at promoters is tightly linked to transcriptional regulation in human cells. At least six different COMPASS-like multi-subunit (SET1/MLL) complexes have been described that contain methyltransferase activity towards H3K4, but a comprehensive and quantitative analysis of these SET1/MLL complexes is lacking. We applied label-free quantitative mass spectrometry to determine the subunit composition and stoichiometry of the human SET1/MLL complexes. Peptides were applied to online nanoLC-MS/MS, using a 120 min acetonitrile gradient (5.6 - 76%). Mass spectra were recorded on an LTQ-Orbitrap-Velos mass spectrometer (Thermo) selecting the 15 most intense precursor ions of every full scan for fragmentation. Raw data were analyzed using MaxQuant 1.3.0.5, with label-free quantification (LFQ), match between runs (between triplicates) and the iBAQ algorithm enabled. MaxQuant default settings were used for peptide identification; Enzyme: Trypsin/P, MS tolerance (FTMS): 6 ppm, max missed cleavages: 2, max charge: 7, MS/MS tolerance (ITMS): 0.5 Da, Peptide FDR: 0.01 and Protein FDR: 0.01. Normalized mass spectrometric intensities (LFQ intensities) were compared between the GFP-tagged and control sample, using an adapted permutation-based false discovery rate (FDR) t-test in Perseus (MaxQuant package).

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: A.H. Smits

LAB HEAD: A.H. Smits

PROVIDER: PXD000172 | Pride | 2013-09-13

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20120601_Velos1_PJ_Lina_Menin_GFPip1.raw	Raw
	20120601_Velos1_PJ_Lina_Menin_GFPip2.raw	Raw
	20120601_Velos1_PJ_Lina_Menin_GFPip3.raw	Raw
	20120601_Velos1_PJ_Lina_RBBP5_GFPip1.raw	Raw
	20120601_Velos1_PJ_Lina_RBBP5_GFPip2.raw	Raw

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Quantitative dissection and stoichiometry determination of the human SET1/MLL histone methyltransferase complexes.

van Nuland Rick R Smits Arne H AH Pallaki Paschalina P Jansen Pascal W T C PW Vermeulen Michiel M Timmers H T Marc HT

Molecular and cellular biology 20130318 10

Methylation of lysine 4 on histone H3 (H3K4) at promoters is tightly linked to transcriptional regulation in human cells. At least six different COMPASS-like multisubunit (SET1/MLL) complexes that contain methyltransferase activity for H3K4 have been described, but a comprehensive and quantitative analysis of these SET1/MLL complexes is lacking. We applied label-free quantitative mass spectrometry to determine the subunit composition and stoichiometry of the human SET1/MLL complexes. We identifi ...[more]

PMID: 23508102

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