Université Paris Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Gif-sur-Yvette, France.
Methods Mol Biol. 2023;2718:137-150. doi: 10.1007/978-1-0716-3457-8_8.
Global acetylation profiling (GAP) consists of heterologous expression of a given N-acetyltransferase (NAT) in Escherichia coli to assess its specificity. The remarkable sensitivity and robustness of the GAP pipeline relies on the very low frequency of known N-terminal acetylated proteins in E. coli, including their degree of N-terminal acetylation. Using the SILProNAQ mass spectrometry strategy on bacterial protein extracts, GAP permits easy acquisition of both qualitative and quantitative data to decipher the impact of any putative NAT of interest on the N-termini of newly acetylated proteins. This strategy allows rapid determination of the substrate specificity of any NAT.
全局乙酰化谱分析（GAP）包括在大肠杆菌中异源表达给定的 N-乙酰转移酶（NAT），以评估其特异性。GAP 管道的显著灵敏度和稳健性依赖于大肠杆菌中已知 N 端乙酰化蛋白的极低频率，包括其 N 端乙酰化程度。在细菌蛋白提取物上使用 SILProNAQ 质谱策略，GAP 允许轻松获取定性和定量数据，以破译任何感兴趣的假定 NAT 对新乙酰化蛋白 N 末端的影响。该策略可快速确定任何 NAT 的底物特异性。
