Proteomics

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WDR76-SPIN1_inWDR76KO_SCAP-MS_MudPIT

ABSTRACT: Serial Capture Affinity Purification (SCAP) samples analyzed using Multidimentional Protein identification Technology (MudPIT) on Orbitrap Velos Elite. Baits are SNAP-SPIN1 and Halo-WDR76. SNAP purification was performed first, then Halo purification was performed. SCAP was performed using whole cell extract from a stable cell line of 293FRT cells in WDR76 KO background. 3 biological replicates of E1, UB2 and E2 fractions.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Michael P. Washburn  

PROVIDER: MSV000086802 | MassIVE | Wed Feb 03 14:45:00 GMT 2021

REPOSITORIES: MassIVE

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Publications

An integrated structural model of the DNA damage-responsive H3K4me3 binding WDR76:SPIN1 complex with the nucleosome.

Liu Xingyu X   Zhang Ying Y   Wen Zhihui Z   Hao Yan Y   Banks Charles A S CAS   Cesare Joseph J   Bhattacharya Saikat S   Arvindekar Shreyas S   Lange Jeffrey J JJ   Xie Yixuan Y   Garcia Benjamin A BA   Slaughter Brian D BD   Unruh Jay R JR   Viswanath Shruthi S   Florens Laurence L   Workman Jerry L JL   Washburn Michael P MP  

Proceedings of the National Academy of Sciences of the United States of America 20240808 33

Serial capture affinity purification (SCAP) is a powerful method to isolate a specific protein complex. When combined with cross-linking mass spectrometry and computational approaches, one can build an integrated structural model of the isolated complex. Here, we applied SCAP to dissect a subpopulation of WDR76 in complex with SPIN1, a histone reader that recognizes trimethylated histone H3 lysine4 (H3K4me3). In contrast to a previous SCAP analysis of the SPIN1:SPINDOC complex, histones and the  ...[more]

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