Development of high-performance amino acid sequensor of peptides

我们已经开发了一种敏感的方法，用于对氨基酸序列和D/L-肽配置进行同步确定，作为荧光Edman试剂盒和BF-D23-D2的循环化/清除试剂，首先，新荧光Edman试剂盒、PSBD-NCS和MSBD-NCS，对氨基酸三唑啉酮衍生物进行了合成和这些试剂盒被敏感地检测到。我们已经设法找到更多稳定的氨基酸衍生物这些试剂。氨基酸三唑啉酮衍生物的DBD-NCS通过硫唑啉化合物转化为碳青霉醇衍生物，并检测到氟等离子体。第三，我们调查了三唑啉酮转化为硫唑啉酮衍生物的转化反应，并发现了甲基氯酸溶液中的氢氯酸在转化反应中有效地抑制了转化反应中的分离。最后，氟原性Edman试剂，MT-NCS，BDsynthesed。硫代氨基甲酸和氨基甲酸衍生物不具有荧光和MTBD-NCS具有荧光的硫代丹酮衍生物，并将其作为对退化产品的干扰峰值的抑制作用的优先权，从而使MTBD-NCS和BF-D23的组合使用我们对氨基酸序列和D/L配置的敏感确定方法的结合使用。本方法是在本研究中开发的，似乎很容易应用于传统自动测序器。

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.