This dataset accompanies a publication in which we present a strategy for acquisition and effective de novo peptide sequencing of complementary CID and 351 nm ultraviolet photodissociation (UVPD) MS/MS pairs. E. coli whole cell lysate is carbamylated to block lysine side chains, digested with trypsin (now active only at Arg residues), and N-terminally tagged with the chromophore AMCA. Three technical replicates were analyzed using a Thermo Velos Pro dual linear ion trap mass spectrometer coupled to a Coherent 351 nm excimer laser. Each precursor ion is isolated twice with the mass spectrometer switching between CID and UVPD activation modes to obtain a complementary MS/MS pair. We modified our UVnovo de novo sequencing software to automatically learn from and interpret fragmentation spectra from any combination of complementary activation methods, and we used this to analyze the CID/UVPD paired spectra. This performance exceeds that of PEAKS and PepNovo on the CID spectra alone and demonstrates that CID/UPVD brings significant advantages for comprehensive and accurate de novo peptide sequencing.
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: 351 nm UVPD ; ultraviolet photodissociation ; AMCA ; de novo
Principal Investigators: (in alphabetical order) |
Edward Marcotte, University of Texas-Austin, United States Jennifer Brodbelt, University of Texas-Austin, United States |
Submitting User: | ahorton |
Horton AP, Robotham SA, Cannon JR, Holden DD, Marcotte EM, Brodbelt JS.
Comprehensive de novo peptide sequencing from MS/MS pairs generated through complementary collision induced dissociation and 351 nm ultraviolet photodissociation.
Anal. Chem. 2017 Feb 24. doi:10.1021/acs.analchem.7b00130.
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