Determination of phosphorylation sites in peptides and proteins employing a volatile Edman reagent

J Protein Chem. 1997 Jul;16(5):329-34. doi: 10.1023/a:1026368201651.


A manual Edman degradation protocol has been developed that allows the identification of phosphorylation sites in 32P-labeled peptides at the subpicomole level. By using both a volatile reagent, trifluoroethyl isothiocyanate, and volatile buffers, extraction steps are rendered unnecessary and cycle times can be reduced to 45 min. The protocol was employed to identify the site of phosphorylation in phosphoserine- and phosphotyrosine-containing peptides.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Binding Sites
  • Chromatography, Thin Layer
  • Electrophoresis
  • Humans
  • Isothiocyanates
  • Peptides / analysis*
  • Peptides / metabolism
  • Phosphorus Radioisotopes
  • Phosphorylation
  • Proteins / analysis*
  • Proteins / metabolism
  • Thiocyanates / chemistry


  • Isothiocyanates
  • Peptides
  • Phosphorus Radioisotopes
  • Proteins
  • Thiocyanates
  • phenylisothiocyanate