Identification of phosphorylation sites in peptides using a gas-phase sequencer

Yuhuan Wang, Carol J. Fiol, Anna De Paoli-Roach, Alexander W. Bell, Mark A. Hermodson, Peter Roach

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

A simple procedure is described for determining the location of phosphorylation sites in phosphopeptides. The method employs measurement of 32P-labeled inorganic phosphate release during Edman degradation cycles using a gas-phase sequencer. The procedure is based on extracting peptides and inorganic phosphate from portions of the sample filter at strategic cycles in the sequence analysis followed by determination of the relative amounts of phosphate and phosphopeptide. One advantage of this technique is the very high recovery of the phosphate associated with the peptide, 80-97% in this study. In the course of this work, it was also found that phosphoserine residues themselves caused reduced efficiency of the Edman degradation as compared with unesterified serine residues. The present procedure has the merit of being simple and easy to apply.

Original languageEnglish
Pages (from-to)537-547
Number of pages11
JournalAnalytical Biochemistry
Volume174
Issue number2
DOIs
StatePublished - Nov 1 1988

Fingerprint

Phosphorylation
Gases
Phosphates
Phosphopeptides
Peptides
Phosphoserine
Degradation
Serine
Sequence Analysis
Recovery

Keywords

  • phosphorylation sites
  • phosphoserine
  • protein sequencing

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Identification of phosphorylation sites in peptides using a gas-phase sequencer. / Wang, Yuhuan; Fiol, Carol J.; De Paoli-Roach, Anna; Bell, Alexander W.; Hermodson, Mark A.; Roach, Peter.

In: Analytical Biochemistry, Vol. 174, No. 2, 01.11.1988, p. 537-547.

Research output: Contribution to journalArticle

Wang, Yuhuan ; Fiol, Carol J. ; De Paoli-Roach, Anna ; Bell, Alexander W. ; Hermodson, Mark A. ; Roach, Peter. / Identification of phosphorylation sites in peptides using a gas-phase sequencer. In: Analytical Biochemistry. 1988 ; Vol. 174, No. 2. pp. 537-547.
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