Bovine catalase: This is an experiment to use differential cysteine alkylation to map the nature of binding of a cysteine-targeting small molecule OBT (for details of the method, please see the Method Section of the manuscript). The alkylating reagents uses besides the studied drug were iodoacetamide (IAA) and N-ethylmaleimide (NEM). As described in the manuscript the experiment includes three routes of serial alkylation experiments termed OBT, IAA, and NEM. Samples were prepared in triplicate and TMT10-plex reagents were used for labeling as follows: OBT replicate 1 (OBT1), 129c; OBT2, 130n; OBT3, 130c; IAA1, 126; IAA2, 127n; IAA3, 127c; NEM1, 128n; NEM2, 128c; NEM3, 129n. An SPS-MS3 method was used for data acquisition on an Orbitrap Fusion Lumos mass spectrometer.
The RAW file for these data is catalase_diffAlk.RAW.
A differential alkylation experiment was also performed as described above for bovine catalase but using recombinant human KEAP1 as substrate. One set of experiments (OBT, IAA, NEM) was performed. TMT10-plex labeling was performed as follows: OBT, 129n; IAA, 128c; NEM, 128n. An SPS-MS3 method was used for data acquisition on an Orbitrap Fusion mass spectrometer.
The RAW file for these data is RAW_file: keap1_diffAlk.RAW.
[doi:10.25345/C5TT20]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: differential alkylation, characterization of thiol binding drug, TMT, Oritrap Fusion (Lumos), SPS-MS3
Principal Investigators: (in alphabetical order) |
Bakhos Tannous, Massachusetts General Hospital and Harvard Medical School, United States |
Submitting User: | whaas |
Number of Files: | |
Total Size: | |
Spectra: | |
Subscribers: | |
Owner | Reanalyses | |
---|---|---|
Experimental Design | ||
Conditions:
|
||
Biological Replicates:
|
||
Technical Replicates:
|
||
Identification Results | ||
Proteins (Human, Remapped):
|
||
Proteins (Reported):
|
||
Peptides:
|
||
Variant Peptides:
|
||
PSMs:
|
||
Quantification Results | ||
Differential Proteins:
|
||
Quantified Proteins:
|
||
Browse Dataset Files | |
FTP Download Link (click to copy):
|