Improved 18O isotope labeling of protein peptide tag.docVIP

 PAGE \* MERGEFORMAT 12 Improved 18O isotope labeling of protein peptide tag Of: Zhao Yan Lu Tao QIAN Xiao Zhuang Jia Wei Ying million [Abstract] 18O isotope labeling reaction for the two important factors - peptides dispersion and trypsin inactivation, the conditions were marked improvements and optimization of inactivation methods. In adding RapigestTM SF H218O and microwave-assisted solvent heated to @ - casein trypsin digested peptide labeling efficiency was significantly upgraded (18O/16O peak area ratiogt; 99%. tag, the reductive alkylation of trypsin completely inactivated chemically modified so that labeled to improve the stability of peptides, place 6 d backcross reaction does not occur. thyroglobulin standard protein mixture digested labeled peptide mass spectrometry results show that: optimal marker method can rapidly and stably labeled protein digested peptides. [Keywords:] 18O isotope labeling, peptides, proteins Abstract In order to optimize the18O labeling method, two key aspects, peptide dispersion and trypsin deactivation were discussed.The addition of RapigestTM SF in H218O and microwave heating enhanced labeling efficiency of @-casein digested peptides (18O/16O ratiogt; 99%). Chemical modification with tris (2-carboxyethyl) phosphine (TCEP) and iodoacetamide (IAA) resulted in trypsin deactivated completely.No significant back-exchange from 18O to16O was observed after labeling in 6 days.The experiment result with peptide mixture from showed that the improved method could be effectively used to label protein and peptide. Keywords 18O stable isotope labeling, Peptide, Proteome 1 Introduction Quantitative proteomics by mass observed between normal and diseased cells or tissues in the protein expression profile differences, the overall level of large-scale screening and identify disease-related proteins, the pathogenic mechanism and clinical applications of important reference information .18 O isotope labeled quantitative pr