Proteomics

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Lambert_P82_VS4_Chromatin_BioID_APMS_12JUL2014

ABSTRACT: This submission contains the mass spectrometry files for the manuscript by Lambert et al. that describes the use of BioID for chromatin associated protein complexes. A detail comparison to a chromatin optimized AP-MS protocol was also performed. This submission contains 39 IDA files for the following baits: empty 3XFLAG control, GFP-BirA-FLAG control, NLS-BirA-FLAG control, H2B-BirA-FLAG, H3.1-BirA-FLAG, 3XFLAG-MED4, BirA-FLAG-MED4, MED20-3XFLAG, MED20-BirA-FLAG, 3XFLAG-MED23 and BirA-FLAG-MED23. Two biological replicates for every bait were analyzed by the standard Gingras lab BioID protocol and by a protocol optimized for chromatin associated baits (AP-MS). See the README file within "Methods and Protocols" and the accompanying File description. Additional details and files are found on prohits-web.lunenfeld.ca.

INSTRUMENT(S): TripleTOF 5600

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

SUBMITTER: Anne-Claude Gingras 

PROVIDER: MSV000078782 | MassIVE | Thu Jul 17 10:44:00 BST 2014

REPOSITORIES: MassIVE

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Publications

Proximity biotinylation and affinity purification are complementary approaches for the interactome mapping of chromatin-associated protein complexes.

Lambert Jean-Philippe JP   Tucholska Monika M   Go Christopher C   Knight James D R JD   Gingras Anne-Claude AC  

Journal of proteomics 20141002

Mapping protein-protein interactions for chromatin-associated proteins remains challenging. Here we explore the use of BioID, a proximity biotinylation approach in which a mutated biotin ligase (BirA*) is fused to a bait of interest, allowing for the local activation of biotin and subsequent biotinylation of proteins in the bait vicinity. BioID allowed for successful interactome mapping of core histones and members of the mediator complex. We explored the background signal produced by the BioID  ...[more]

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