Mass spectrometry-based methods for protein identification and ：5基于质谱的蛋白质鉴定和方法.pptVIP

MS-based methods for protein identification phosphorylation site analysis ????? ??? Mass Spectrometer(MS) ??? MS ?? ???? ??? ????? ??? ? ?? ???(fragments)? ????. ??? ???? ??? ??/??(m/z)?? ?? ????? ???? ???? ???? signal? ???. ???? ??/???? ???? ??? ?(Abundance)? ??? ???? mass spectrum? ????, ? mass spectrum? ???? ????? ????? ? ? ??. ??? ???? ??? ?? ?? MS component Sample inlet ??????? ??? MS?? ????? ???? ??? ?? Ion source ????? ?????? ? ?? ???? ???. ??? ???? MS analyzer ??? ????? Mass analyzer ???? m/z ratio? ?? ????? ????? Ion detector ?? ??? ? ?? ???? ???? ???? ??, ???? signal? ???? Vacuum system MS ? ????? 10-4 ~ 10-9 Torr ? ??? ?? ??? ??? ??? ??? ? ??? ?? Data System MS ? ? ?????? ??? ???? ????? ??? ?????? ? ? ?? ??? Ion source Gaseous sample introduction - EI(electron ionization) - CI(chemical Ionization) Liquid sample introduction - FAB(fast atom bombardment) - ESI(electrospray ionization)(soft ionization) Solid sample introduction - MALDI(soft ionization) (matrix-assisted laser desorption/ionization) EI(electron ionization) ??? ????? ?? filament? ???? (+) ??? ??? ??? ??? ???? ??? ? ?? ??? ???? sample? ??? ???? ???? ?? ?? ??? ?? ???? M+? ?? ? ??? ? ???? ???? ??? ??? spectrum? ??? ?? ??? ?? ??? M + e- ? M+ + 2e- CI(chemical Ionization) ??? filament?? ??, ??? ??? 106 ??? ?? reagent gas? ???? ??? ?????? ? reagent gas ion? sample gas? ?? ? sample gas? fragmentation ??? ?? ??? reagent gas ion ? complex? ???? ??. ?? ?? ???? ???? ??? EI ?? ??? ?? ?? ?? ?? ??? ??? ?? ??? ??? ??? ?? ???. R(CH4) + e- ? R+ + 2 e- R+ + M ? M1+ + N1 M1+ ? M2+ + N2 MALDI process matrix? sample? ? 1000~10,000 : 1? ?? ??? ??? ??(acidic organic solvent- TFA+MeCN)? ???? ? ???? ??? probe ? ?? ?? ????? ??? ?? ????? ???? ??? matrix ? ?? ???? ??? ?? ? ? ?? ???(UV at 337nm, IR at 2.94um) ? sample-matrix ??? ???? ???? matrix? ?? sample? ???? ?? ???? ????. ???? protonation/deprotonation, cation attachment/cation detach