Histone Sample Preparation for Bottom-Up Mass Spectrometry: A Roadmap to Informed Decisions.
GingisREX
coverage
epigenetics
histone code
mass spectrometry
sample preparation
workflow optimization
Journal
Proteomes
ISSN: 2227-7382
Titre abrégé: Proteomes
Pays: Switzerland
ID NLM: 101621966
Informations de publication
Date de publication:
21 Apr 2021
21 Apr 2021
Historique:
received:
26
02
2021
revised:
08
04
2021
accepted:
19
04
2021
entrez:
30
4
2021
pubmed:
1
5
2021
medline:
1
5
2021
Statut:
epublish
Résumé
Histone-based chromatin organization enabled eukaryotic genome complexity. This epigenetic control mechanism allowed for the differentiation of stable gene-expression and thus the very existence of multicellular organisms. This existential role in biology makes histones one of the most complexly modified molecules in the biotic world, which makes these key regulators notoriously hard to analyze. We here provide a roadmap to enable fast and informed selection of a bottom-up mass spectrometry sample preparation protocol that matches a specific research question. We therefore propose a two-step assessment procedure: (i) visualization of the coverage that is attained for a given workflow and (ii) direct alignment between runs to assess potential pitfalls at the ion level. To illustrate the applicability, we compare four different sample preparation protocols while adding a new enzyme to the toolbox, i.e., RgpB (GingisREX
Identifiants
pubmed: 33919160
pii: proteomes9020017
doi: 10.3390/proteomes9020017
pmc: PMC8167631
pii:
doi:
Types de publication
Journal Article
Langues
eng
Subventions
Organisme : Fonds Wetenschappelijk Onderzoek
ID : 11B4518N
Organisme : Fonds Wetenschappelijk Onderzoek
ID : 12E9716N
Organisme : Fonds Wetenschappelijk Onderzoek
ID : 3S031319
Organisme : Bijzonder Onderzoeksfonds UGent
ID : 01D22020
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