Journal
PROTEOMICS
Volume 14, Issue 10, Pages 1130-1140Publisher
WILEY
DOI: 10.1002/pmic.201300438
Keywords
Human histone H4; PTM analysis; Technology; Top-down proteomics
Funding
- NIH [R01GM067193, 1R01NS065263]
- NSF [CHE-1012622, ABI-1062432, DMR-11-57490]
- Welch Foundation [F-1155]
- Genome BC for Science and Technology Innovation Centre
- Department of Energy's Office of Biological and Environmental Research
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Pilot Project #1the identification and characterization of human histone H4 proteoforms by top-down MSis the first project launched by the Consortium for Top-Down Proteomics (CTDP) to refine and validate top-down MS. Within the initial results from seven participating laboratories, all reported the probability-based identification of human histone H4 (UniProt accession P62805) with expectation values ranging from 10(-13) to 10(-105). Regarding characterization, a total of 74 proteoforms were reported, with 21 done so unambiguously; one new PTM, K79ac, was identified. Inter-laboratory comparison reveals aspects of the results that are consistent, such as the localization of individual PTMs and binary combinations, while other aspects are more variable, such as the accurate characterization of low-abundance proteoforms harboring >2 PTMs. An open-access tool and discussion of proteoform scoring are included, along with a description of general challenges that lie ahead including improved proteoform separations prior to mass spectrometric analysis, better instrumentation performance, and software development.
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