Development of high-performance amino acid sequensor of peptides

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We have developed a sensitive method for the simultaneous determination of an amino acid sequence and D/L-configuration of peptides with fluorogenic Edman reagents and BFィイD23ィエD2 as cyclization/cleavage reagent.First, New fluorogenic Edman reagents, PSBD-NCS and MSBD-NCS, were synthesized and the amino acids thiazolinone derivatives of these reagents were sensitively detected.Secondly, we tried to detect more stable amino acids derivatives of these reagents. The amino acids thiazolininone derivatives of DBD-NCS were converted to carbamoyl derivatives via thiocarbamoyl compounds and detected fluorimetrically.Thirdly, we investigated the conversion reaction of thiazolinone to thiohydantoin derivatives and found out that hydrochloric acid in methanol solution efficiently suppressed the racemization in the conversion reaction.Finally, fluorogenic Edman reagent, MTBD-NCS, was synthesized. The thiocamamoyl and carbamoyl derivatives were not fluorescent and thiohydantoin derivatives of MTBD-NCS were fluorescent, giving the superiority with regard to avoidance from the interferring peaks of the degradation products.The combined use of MTBD-NCS and BFィイD23ィエD2 gave us the sensitive determination method of amino acid sequence and D/L-configuration of peptides. This method developed in this study seems to be easily applicable to the conventional automatic sequensor.