Dataset Information

P65-HaloTag copurifying proteins in human 293T cells

ABSTRACT: In effort to develop methodology for targeted top down mass spectrometry of NF kappa B p65 from human cells, we evaluated the utility of HaloTag for purification and analysis of recombinant protein. During our study, two datasets of bottom up LC-MS/MS were generated: one from in-gel digestion of the predominant band following p65-HaloTag purification, another from in-solution digestion of all the proteins present in a p65-HaloTag purification. p65-HaloTag copurifying proteins identified in our datasets include the known interactors c-Rel, NF-kappaB p105, NF-kappaB p100, and NF-kappaB inhibitor beta. Over 100 proteins were identified by at least two peptides using a Mascot ion cut-off score of 30.

INSTRUMENT(S): LTQ FT, LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

SUBMITTER: John Savaryn

LAB HEAD: Neil Kelleher

PROVIDER: PXD001932 | Pride | 2018-10-19

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20140703_B1major.RAW	Raw
	20140703_B1major_2.RAW	Raw
	20140703_B1major_2_ITMS_CID.mgf	Mgf
	20140703_B1major_2_ITMS_CID.pride.mgf.gz	Mgf
	20140703_B1major_ITMS_CID.mgf	Mgf

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1 - 5 of 15

Publications

Targeted analysis of recombinant NF kappa B (RelA/p65) by denaturing and native top down mass spectrometry.

Savaryn John Paul JP Skinner Owen S OS Fornelli Luca L Fellers Ryan T RT Compton Philip D PD Terhune Scott S SS Abecassis Mike M MM Kelleher Neil L NL

Journal of proteomics 20150504

Measuring post-translational modifications on transcription factors by targeted mass spectrometry is hampered by low protein abundance and inefficient isolation. Here, we utilized HaloTag technology to overcome these limitations and evaluate various top down mass spectrometry approaches for measuring NF-κB p65 proteoforms isolated from human cells. We show isotopic resolution of N-terminally acetylated p65 and determined it is the most abundant proteoform expressed following transfection in 293T ...[more]

PMID: 25952688

Dataset Information

P65-HaloTag copurifying proteins in human 293T cells

Dataset's files

Publications

Targeted analysis of recombinant NF kappa B (RelA/p65) by denaturing and native top down mass spectrometry.

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