通过将体内稳定同位素标记细胞培养技术(AACT/SILAC)与LC-MS/MS联用,对哺乳动物细胞中TNF-α/NF-κB信号通路相关蛋白在TNF-α及TSA存在情况下乙酰化变化情况进行了研究.在TSA协同处理的、TNF-α刺激的细胞中鉴定到3个乙酰化蛋白,其中核糖体蛋白L4(60SRibosomal Protein L4,RPL4)已被报道存在乙酰化的后修饰,转铁蛋白受体1(Transferrin Receptor Protein 1,TfR1)和凋亡诱导因子3(ApoptosisInducing Factor,Mitochondrion-associated,3,AIFM3)尚未见报道.
A novel detection method for quaternization of amino acid and peptide was reported. In this method Ag 2CO 3 was employed instead of KHCO 3 to catalyze the occurrence of quaternization reaction. The absence of metal ions in the reaction system enabled us to in situ detect the quaternizing products by using electrospray ionization mass spectrometry(ESI-MS). We analyzed quaternizing products of six amino acids and two peptides in situ, and obtained the ratio of major product to by-product, which suggests the new method possesses prominent advantages over the old one. This method could offer a potential chance to combine quaternization with mass spectrometry which is helpful for the future use of quaternization reaction in quantitative proteomics.
