title: An MRM-based workflow for absolute quantitation of lysine-acetylated metabolic enzymes in mouse liver.
abstract: As a key post-translational modification mechanism, protein acetylation plays critical roles in regulating and/or coordinating cell metabolism. Acetylation is a prevalent modification process in enzymes. Protein acetylation modification occurs in sub-stoichiometric amounts; therefore extracting biologically meaningful information from these acetylation sites requires an adaptable, sensitive, specific, and robust method for their quantification. In this work, we combine immunoassays and multiple reaction monitoring-mass spectrometry (MRM-MS) technology to develop an absolute quantification for acetylation modification. With this hybrid method, we quantified the acetylation level of metabolic enzymes, which could demonstrate the regulatory mechanisms of the studied enzymes. The development of this quantitative workflow is a pivotal step for advancing our knowledge and understanding of the regulatory effects of protein acetylation in physiology and pathophysiology.
authors.affiliation: Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, 201199, Shanghai, China. hongxiuyu@fudan.edu.cn pyyang@fudan.edu.cn.
authors.forename: Leilei
authors.initials: L
authors.lastname: Xu
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.forename: Cuiping
authors.initials: C
authors.lastname: Zhang
authors.forename: Xue
authors.initials: X
authors.lastname: Qin
authors.forename: Hongxiu
authors.initials: H
authors.lastname: Yu
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
mesh_terms: D000107:Acetylation; D000818:Animals; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D004798:Enzymes / Q000032:analysis* / Q000378:metabolism; D055786:Gene Knockout Techniques; D008099:Liver / Q000201:enzymology*; D008239:Lysine / Q000737:chemistry*; D051379:Mice; D010455:Peptides / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D004798:Enzymes; D010455:Peptides; D008239:Lysine