An Improved Top-Down Mass Spectrometry Characterization of Chlamydomonas reinhardtii Histones and Their Post-translational Modifications
Journal Article
·
· Journal of the American Society for Mass Spectrometry
- Donald Danforth Plant Science Center, St. Louis, MO (United States); Washington Univ., St. Louis, MO (United States)
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Saint Mary’s College of California, Moraga, CA (United States)
- Donald Danforth Plant Science Center, St. Louis, MO (United States)
We present an updated analysis of the linker and core histone proteins and their proteoforms in the green microalga Chlamydomonas reinhardtii by top-down mass spectrometry (TDMS). The combination of high-resolution liquid chromatographic separation, robust fragmentation high, mass spectral resolution, the application of a custom search algorithm, and extensive manual analysis enabled the characterization of 86 proteoforms across all four core histones H2A, H2B, H3 and H4, and the linker histone H1. All canonical H2A paralogs, which vary in their C-termini, were identified, along with an unreported non-canonical variant that had both high levels of acetylation and C-terminal truncations. Similarly, a majority of the canonical H2B paralogs were identified, along with a smaller non-canonical variant, H2B.v, that was highly acetylated. Histone H4 exhibited a novel acetylation profile that differs significantly from that found in other organisms. A majority of H3 was monomethylated at K4, while a small fraction of H3 was trimethylated at K4 and also highly acetylated.
- Research Organization:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- National Institutes of Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC), Biological and Environmental Research (BER)
- Grant/Contract Number:
- AC05-76RL01830
- OSTI ID:
- 1808068
- Report Number(s):
- PNNL-SA--160023
- Journal Information:
- Journal of the American Society for Mass Spectrometry, Journal Name: Journal of the American Society for Mass Spectrometry Journal Issue: 7 Vol. 32; ISSN 1044-0305
- Publisher:
- American Society for Mass SpectrometryCopyright Statement
- Country of Publication:
- United States
- Language:
- English
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