期刊
ANALYTICAL CHEMISTRY
卷 91, 期 19, 页码 12336-12343出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.9b02656
关键词
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资金
- Deutsche Forschungsgemeinschaft (DFG, Germany) Research Training Group 2155, ProMoAge
- Federal Government of Germany
- State of Thuringia
Recently discovered acylation by reactive acyl-CoA species is considered a novel regulatory mechanism in epigenetics and metabolism. Established analytical methods like Western blotting and proteomics fail to detect the plethora of acylation structures in a single analysis and lack the ability of absolute quantitation. In this paper, we developed an HPLC-MS/MS method for the simultaneous detection and quantitation of 14 acylated lysine species in biological samples. Extensive effort was invested into method validation resulting in recovery rates between 75 and 93% and levels of detection in the nanomolar range. Thus, we were able to quantitate 8 acylation structures in mouse liver, kidney, heart, and brain. Further enrichment by repetitive HPLC fractionation resulted in the quantitation of 6 additional acylation structures including 4 novel modifications: N-6-acetoacetyl lysine, N-6-isovaleryl lysine, N-6-(2-methylbutyryl) lysine, and N-6-tiglyl lysine.
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