Near-infrared fluorescence probes for enzymes based on binding affinity modulation of squarylium dye scaffold

Daihi Oushiki, Hirotatsu Kojima, Yuki Takahashi, Toru Komatsu, Takuya Terai, Kenjiro Hanaoka, Makiya Nishikawa, Yoshinobu Takakura, Tetsuo Nagano

研究成果: Article査読

51 被引用数 (Scopus)

抄録

We present a novel design strategy for near-infrared (NIR) fluorescence probes utilizing dye-protein interaction as a trigger for fluorescence enhancement. The design principle involves modification of a polymethine dye with cleavable functional groups that reduce the dye's protein-binding affinity. When these functional groups are removed by specific interaction with the target enzymes, the dye's protein affinity is restored, protein binding occurs, and the dye's fluorescence is strongly enhanced. To validate this strategy, we first designed and synthesized an alkaline phosphatase (ALP) sensor by introducing phosphate into the squarylium dye scaffold; this sensor was able to detect ALP-labeled secondary antibodies in Western blotting analysis. Second, we synthesized a probe for β-galactosidase (widely used as a reporter of gene expression) by means of β-galactosyl substitution of the squarylium scaffold; this sensor was able to visualize β-galactosidase activity both in vitro and in vivo. Our strategy should be applicable to obtain NIR fluorescence probes for a wide range of target enzymes.

本文言語English
ページ(範囲)4404-4410
ページ数7
ジャーナルAnalytical Chemistry
84
10
DOI
出版ステータスPublished - 2012 5月 15
外部発表はい

ASJC Scopus subject areas

  • 分析化学

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