Improved molecular weight-based processing of intact proteins for interrogation by quadrupole-enhanced FT MS/MS

Yi Du, Fanyu Meng, Steven M. Patrie, Leah M. Miller, Neil L. Kelleher

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Complete coverage of protein primary structure is demonstrated for 37 yeast protein forms between 6 and 30 kDa in an improved platform for Top Down mass spectrometry (MS). Tandem mass spectrometry (MS/MS) for protein identification with 100% sequence coverage is achieved in a highly automated fashion with 15-300-fold less sample amounts than an initial report of a proteome fractionation approach employing preparative gel electrophoresis with an acid-labile surfactant to facilitate reversed phase separation in a second dimension. Using a quadrupole-enhanced Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FTICRMS) improves the dynamic range for protein detection by ∼50-fold and MS/MS by ∼30-fold. The technology development illustrated here typifies an accelerating effort to detect whole proteins in a more general and higher throughput fashion for improved biomarker identification and detection of diverse post-translational modifications. Capillary RPLC is used in both off-line and on-line modes, with one on-line LC/FTMS sample providing 25 observed protein forms from 11 to 22 kDa.

Original languageEnglish (US)
Pages (from-to)801-806
Number of pages6
JournalJournal of Proteome Research
Volume3
Issue number4
DOIs
StatePublished - Jul 1 2004

Keywords

  • Capillary RPLC
  • Miniaturization
  • Q-FTMS
  • Top down proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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