Mapping of protein phosphorylation by dual enzyme digestion and MALDI/quadrupole orthogonal TOF mass spectrometry

Jun Han, Marshall Pope, Christoph Borchers, Lee M. Graves

Research output: Contribution to conferencePaperpeer-review

Abstract

The use of sequential trypsin and proteinase K proteolytic digestion, immobilized metal affinity chromatography (IMAC) and MALDI-qTOFMS for the characterization of protein phosphorylation was discussed. Its utility was tested on a synthetic phosphopeptide and two standard phosphoproteins. It was demonstrated that nonspecific proteinase K could be combined with tryptic digestion and specific IMAC purification to effectively reduce large phosphopeptides to suitable size, which favored MS/MS sequencing and improved the elucidation of protein phosphorylation by MALDI/qTOFMS. It was also found that the process could be used as a supplementary method to facilitate the analysis of large tryptic phosphopeptides that cannot be directly sequenced by MS/MS.

Original languageEnglish
Pages559-560
Number of pages2
Publication statusPublished - 2002
Externally publishedYes
EventProceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics - Orlando, FL, United States
Duration: 2 Jun 20026 Jun 2002

Conference

ConferenceProceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics
Country/TerritoryUnited States
CityOrlando, FL
Period2/06/026/06/02

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