Journal
BULLETIN OF THE KOREAN CHEMICAL SOCIETY
Volume 33, Issue 10, Pages 3233-3240Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.5012/bkcs.2012.33.10.3233
Keywords
Electron transfer Dissociation (ETD); Collisionally activated dissociation (CAD); Trypsin-immobilization; Immobilized enzyme reactor (IMER); Monolithic reactor
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Funding
- Korea Basic Science Institute [T32608]
- National Research Foundation of Korea (NRF)
- Ministry of Education, Science and Technology [2012R1A1A2006532]
- National Research Foundation of Korea [2012R1A1A2006532] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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We demonstrated the combined applications of online protein digestion using trypsin immobilized enzyme reactor (IMER) and dual tandem mass spectrometry with collisionally activated dissociation (CAD) and electron transfer dissociation (ETD) for tryptic peptides eluted through the trypsin-IMER. For the trypsin-IMER, the organic and inorganic hybrid monolithic material was used. By employing the trypsin-IMER, the long digestion time could be saved with little or no sacrifice of the digestion efficiency, which was demonstrated for standard protein samples. For three model proteins (cytochrome c, carbonic anhydrase, and bovine serum albumin), the tryptic peptides digested by the IMER were analyzed using LC-MS/MS with the dual application of CAD and ETD. As previously shown by others, the dual application of CAD and ETD increased the sequence coverage in comparison with CAD application only. In particular, ETD was very useful for the analysis of highly-protontated peptide cations, e.g., >= 3+. The combination approach provided the advantages of both trypsin-IMER and CAD/ETD dual tandem mass spectrometry applications, which are rapid digestion (i.e., 10 min), good digestion efficiency, online coupling of trypsin-IMER and liquid chromatography, and high sequence coverage.
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