Simple strategies to enhance discovery of acetylation post-translational modifications by quadrupole-orbitrap LC-MS/MS

被引:2
作者
Manning, Andrew J. [1 ]
Lee, Jiyoung [1 ,2 ]
Wolfgeher, Donald J. [1 ]
Kron, Stephen J. [1 ]
Greenberg, Jean T. [1 ]
机构
[1] Univ Chicago, Dept Mol Genet & Cell Biol, 920 E 58Th St, Chicago, IL 60637 USA
[2] Korean Res Inst Biosci & Biotechnol, Jeongeup, South Korea
来源
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS | 2018年 / 1866卷 / 02期
基金
美国国家科学基金会;
关键词
Post-translational modification; Plant PTMs; Acetylation; C-13; O-acetylation; Q-exactive hybrid quadrupole-orbitrap; TANDEM MASS-SPECTROMETRY; PROTEIN IDENTIFICATION; LYSINE ACETYLATION; SITE LOCALIZATION; MODIFIED PEPTIDES; MANUAL VALIDATION; EFFECTOR HOPZ1A; PHOSPHORYLATION; SPECTRA; QUANTIFICATION;
D O I
10.1016/j.bbapap.2017.10.006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Enzyme-dependent post-translational modifications (PTMs) mediate the cellular regulation of proteins and can be discovered using proteomics. However, even where the peptides of interest can be enriched for analysis with state-of-the-art LC-MS/MS tools and informatics, only a fraction of peptide ions can be identified confidently. Thus, many PTM sites remain undiscovered and unconfirmed. In this minireview, we use a case study to discuss how the use of inclusion lists, turning off isotopic exclusion, and manual validation significantly increased depth of coverage, facilitating discovery of acetylation sites in targets of an acetyltransferase virulence factor. These underutilized strategies have the potential to help answer many mechanistic biological questions that large-scale proteomic studies cannot.
引用
收藏
页码:224 / 229
页数:6
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