Hasina Yasmin, Takayuki Shibata, Mohammed Shafikur Rahman, Tsutomu Kabashima, Masaaki Kai
Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Japan.
Analytica chimica acta 2012 Apr 6A novel fluorescence (FL) reaction for N-terminal Gly-containing peptides has been developed using 3,4-dihydroxyphenylacetic acid (3,4-DHPAA). The reaction of the peptides with 3,4-DHPAA was carried out in borate buffer (pH 8.0) in the presence of sodium periodate at 37°C for 10 min, and the FL was measured with a spectrofluorimeter at the excitation and emission wavelengths of 370 nm and 465 nm, respectively. The 3,4-DHPAA reagent generated particularly strong FL for peptides containing Gly at their N-termini. When various other bio-substances, such as amino acids, sugars, nucleic bases, nucleotides, and proteins, were reacted with 3,4-DHPAA, no FL was observed. Under optimized reaction conditions, the lower detection limit of 0.25 μmol L(-1) was obtained for the N-terminal Gly-containing peptides of Gly-Pro (GP) and Gly-Pro-Pro (GPP), which gave 3 times greater FL intensity than that observed for the reagent blank. The proposed reaction with 3,4-DHPAA as a fluorogenic reagent is selective and sensitive for the detection of N-terminal Gly-containing peptides, and therefore, this method could be a useful tool for the determination of these particular oligopeptides. Copyright © 2012 Elsevier B.V. All rights reserved.
Hasina Yasmin, Takayuki Shibata, Mohammed Shafikur Rahman, Tsutomu Kabashima, Masaaki Kai. Selective and sensitive determination of peptides using 3,4-dihydroxyphenylacetic acid as a fluorogenic reagent. Analytica chimica acta. 2012 Apr 6;721:162-6
PMID: 22405315
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