Synthetic Assembly of Bifluorescence-Labeled Glycopolymers as Substrates for Assaying 伪-Amylase by Resonance Energy Transfer

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• 作者：Koji Matsuoka ; Hirokatsu Arai ; Hiroyuki Oka ; Tetsuo Koyama ; Ken Hatano
• 刊名：ACS Macro Letters
• 出版年：2012
• 出版时间：February 21, 2012
• 年：2012
• 卷：1
• 期：2
• 页码：266-269
• 全文大小：285K
• 年卷期：v.1,no.2(February 21, 2012)
• ISSN：2161-1653

文摘

To meet the need for a convenient substrate for sensitive and continuous assay for 伪-amylase, we developed a fluorescence resonance energy transfer (FRET)-based polymer substrate. Radical copolymerization of FRET-component monomers in different ratios of fluorogenic donor and acceptor was utilized to prepare such polymers. A glycomonomer as a fluorogenic donor was derived from naphthylmethylated maltotetraose, and a dansyl derivative monomer was used as an acceptor. Their mixture and acryl amide were copolymerized in a typical radical polymerization to yield a bifluorescence-labeled polymer in good yield. All of the polymers showed effective FRET and were used for the continuous assay of human salivary 伪-amylase. The time course of 伪-amylase reactions led to the apparent kinetic parameters of K_m = 4 渭M and V_max = 0.29 nmol/min. The results strongly suggested that FRET-sensitive polymers are conveniently accessible and applicable for the sensitive determination of biochemical events.