I have a a tripeptide which was isolated using prep LC. I have determined its accurate mass and empirical formula using LC-QTOf-MS. I went further and did acid digestion which has enabled determination of individual amino acids. Any suggestions on the simplest Edman degradation protocol I can follow to elucidate the structure? Thanks
try this
• Coupling of the phenylisothiocyanate (PITC, Edman reagent) to the alpha-amine of the polypeptide chain under basic conditions to form a phenylthiocarbamyl (PTC) moiety.
• Cleavage under mild acidic conditions generate a free amino terminus on the polypeptide and
an anilinothiazolinone (ATZ) adducted amino acid.
• The latter is extracted and further converted to a more stable phenylthiohydantoin (PTH)
derivative. The resulting PTH residue is analyzed by HPLC and retention times compared to
that of standards PTH amino acids.
try to see this Cheseto, https://www.amherst.edu/system/files/media/1867/lab%2520manual%2520A%252009.pdf
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