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).
[doi:10.25345/C53F66]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: PROTAC, CBP, p300, human cell lines, TMT11, SPS-MS3, Orbitrap Fusion, Orbitrap Lumos
Principal Investigators: (in alphabetical order) |
Christopher J. Ott, Massachusetts General Hospital Cancer Center and Harvard Medical School, United States |
Submitting User: | whaas |
Vannam R, Sayilgan J, Ojeda S, Karakyriakou B, Hu E, Kreuzer J, Morris R, Herrera Lopez XI, Rai S, Haas W, Lawrence M, Ott CJ.
Targeted degradation of the enhancer lysine acetyltransferases CBP and p300.
Cell Chem Biol. 2021 Apr 15;28(4):503-514.e12. Epub 2021 Jan 4.
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