RRC ID 43772
著者 Oushiki D, Kojima H, Takahashi Y, Komatsu T, Terai T, Hanaoka K, Nishikawa M, Takakura Y, Nagano T.
タイトル Near-infrared fluorescence probes for enzymes based on binding affinity modulation of squarylium dye scaffold.
ジャーナル Anal Chem
Abstract 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.
巻・号 84(10)
ページ 4404-10
公開日 2012-5-15
DOI 10.1021/ac300061a
PMID 22482531
MeSH Alkaline Phosphatase / chemistry Alkaline Phosphatase / metabolism* Animals Fluorescent Dyes / chemistry* HEK293 Cells Humans Mice Protein Binding Quantum Theory Spectroscopy, Near-Infrared* beta-Galactosidase / chemistry beta-Galactosidase / metabolism*
IF 6.785
引用数 32
WOS 分野 CHEMISTRY, ANALYTICAL
リソース情報
ヒト・動物細胞