| 摘要 |
A method for chemically generating a set of N-terminally truncated peptides (ladder), suitable for analysis by mass spectrometry (MS), from a peptide or protein using carbon disulfide (CS2). The method consists of: 1) a coupling step employing CS2, a tert-amine catalyst, and a solvent under conditions that maximize the yield of dithiocarbamyl-peptide, and 2) a cleavage step employing an acid, which catalyzes two alternate pathways, removal of the N-terminal residue and regeneration of the starting peptide, with no other modification. The simultaneous operation of both pathways generates a ladder without further intervention. This pair of steps may be repeated a controlled but unlimited number of times with drying between each step in order to generate a ladder of any desired length. Finally, the processed peptide is analyzed by MS, wherein the sequence of the peptide is deduced from the mass ladder. The ratio of cleavage to regeneration permits the distinction of isoleucine and leucine.
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