Development of an Amino Acid Sequence and D/L-Configuration Determination Method of Peptide with a New Fluorescence Edman Reagent, 7-Methylthio-4-(2,1,3-b
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- 作者：Akira Toriba ; Kentaro Adzuma ; Tomofumi Santa ; and Kazuhiro Imai
- 刊名：Analytical Chemistry
- 出版年：2000
- 出版时间：February 15, 2000
- 年：2000
- 卷：72
- 期：4
- 页码：732 - 739
- 全文大小：104K
- 年卷期：v.72,no.4(February 15, 2000)
- ISSN：1520-6882
文摘
On the basis of the relationship between the fluorescencecharacteristics of the benzofurazan compounds and theHammett constants (
p), a new fluorescence Edmanreagent, 7-methylthio-4-(2,1,3-benzoxadiazolyl) isothiocyanate (MTBD-NCS) was designed and synthesized.MTBD-thiohydantoin (TH)-amino acid derivatives produced by the Edman sequencing method gave fluorescence, whereas other degradation byproducts such asMTBD-thiocarbamoyl (TC)- or carbamoyl (CA)-aminoacids did not fluoresce. MTBD-NCS was applicable as anEdman sequencing reagent to the simultaneous determination of both the sequence and D/L-configuration ofamino acids in peptides. Boron trifluoride (BF3) and HCl/methanol were adopted as the cyclization/cleavage andconversion reagents to suppress the amino acid residueracemization. The MTBD-TH-amino acids were separatedon a reversed-phase column for amino acid sequencing,and their enantiomers were resolved on two types ofpolysaccharide-based chiral stationary phases for D/L-configuration determination. The method was successfullyapplied to the sequence and D/L-configuration determination of D-amino acid-containing peptide [D-Ala2]-deltorphin II.
p), a new fluorescence Edmanreagent, 7-methylthio-4-(2,1,3-benzoxadiazolyl) isothiocyanate (MTBD-NCS) was designed and synthesized.MTBD-thiohydantoin (TH)-amino acid derivatives produced by the Edman sequencing method gave fluorescence, whereas other degradation byproducts such asMTBD-thiocarbamoyl (TC)- or carbamoyl (CA)-aminoacids did not fluoresce. MTBD-NCS was applicable as anEdman sequencing reagent to the simultaneous determination of both the sequence and D/L-configuration ofamino acids in peptides. Boron trifluoride (BF3) and HCl/methanol were adopted as the cyclization/cleavage andconversion reagents to suppress the amino acid residueracemization. The MTBD-TH-amino acids were separatedon a reversed-phase column for amino acid sequencing,and their enantiomers were resolved on two types ofpolysaccharide-based chiral stationary phases for D/L-configuration determination. The method was successfullyapplied to the sequence and D/L-configuration determination of D-amino acid-containing peptide [D-Ala2]-deltorphin II.