Quantitative determination of phosphoserine by high-performance liquid chromatography as the phenylthiocarbamyl-s-ethylcysteine : Application to picomolar amounts of peptides and proteins

✍ Scribed by Helmut E. Meyer; Kristine Swiderek; Edeltraut Hoffmann-Posorske; Horst Korte; Ludwig M.G. Heilmeyer Jr.

• Publisher: Elsevier Science
• Year: 1987
• Tongue: English
• Weight: 497 KB
• Volume: 397
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(01)84994-3

✦ Synopsis

A method is described that permits the phosphoserine content of proteins and peptides to be determined in picomolar amounts. A micro-batch reaction first converts phosphoserine into S-ethylcysteine. Hydrolysis with 6 M hydrochloric acid then yields the free amino acid, which is coupled with phenyl isothiocyanate to give the corresponding phenylthiocarbamylamino acid. This derivative is determined quantitatively in the range 10-20 pmol by reversed-phase high-performance liquid chromatography. The method works well with either small peptides or proteins in the low picomole range.