Targeted degradation of the enhancer lysine acetyltransferases CBP and p300
Ontology highlight
ABSTRACT: The enhancer factors CBP/p300 maintain gene expression programs through lysine acetylation of chromatin and transcriptional regulators and by scaffolding functions mediated by several protein-protein interaction domains. We have designed dCBP-1, a compound that induces degradation of CBP/p300, and profiled its activity inducing global proteome changes in HAP1 and MM1S cells. The proteome changes were mapped using multiplexed quantitative proteomics (TMT11) and applying the SPS-MS3 method on either an Orbitrap Fusion or an Orbitrap Lumos Fusion mass spectrometer. The proteome was mapped 3 hours and 6 hours upon drug treatment. Samples were covered over two TMT sets for both time points. Samples for the 3 hours time points (HAP1 and MM1S) were analyzed using a total of 48 fractions on an Orbitrap Lumos mass spectrometer. Samples for the 6 hours time point (MM1S) were analyzed using a total of 20 fractions on an Orbitrap Fusion mass spectrometer. Samples were analyzed in three replicates (rep1-rep3). Pooled tryptic digests were used as bridge channels to compare proteome profiles from the two TMT sets mapped per time point.
Samples were labeled as described in the following:
3 hours:
HAP1_DMSO_3h_rep1 (Ott_3hours_TMT1_129c);
HAP1_DMSO_3h_rep2 (Ott_3hours_TMT1_130n);
HAP1_DMSO_3h_rep3 (Ott_3hours_TMT1_130c);
HAP1_dCBP1_3h_rep1 (Ott_3hours_TMT2_129c);
HAP1_dCBP1_3h_rep2 (Ott_3hours_TMT2_130n);
HAP1_dCBP1_3h_rep3 (Ott_3hours_TMT2_130c);
MM1S_DMSO_3h_rep1 (Ott_3hours_TMT1_126);
MM1S_DMSO_3h_rep2 (Ott_3hours_TMT1_127n);
MM1S_DMSO_3h_rep3 (Ott_3hours_TMT1_127c);
MM1S_dCBP1_3h_rep1 (Ott_3hours_TMT1_128n);
MM1S_dCBP1_3h_rep2 (Ott_3hours_TMT1_128c);
MM1S_dCBP1_3h_rep3 (Ott_3hours_TMT1_129n);
Bridge channels (Ott_3hours_TMT1 and TMT2, 131n).
6 hours:
TMT1_128c (Ott_6hours_TMT1_128c);
TMT2_126 (Ott_6hours_TMT2_126);
TMT2_129c (Ott_6hours_TMT2_129c);
TMT1_129n (Ott_6hours_TMT1_129n);
TMT2_127n (Ott_6hours_TMT2_127n);
TMT2_130n (Ott_6hours_TMT2_130n);
Bridge channels (Ott_6hours_TMT1 and TMT2, 131c).
INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Fusion
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: 
Christopher J. Ott
PROVIDER: MSV000086457 | MassIVE | Sun Nov 15 08:42:00 GMT 2020
REPOSITORIES: MassIVE
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