固体発光性色素を用いたシアリダーゼライブイメージングプローブの開発

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  • Development of Sialidase Live-imaging Probe Using a Solid Fluorescent Pigment Dye
  • コタイ ハッコウセイ シキソ オ モチイタ シアリダーゼライブイメージングプローブ ノ カイハツ

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<p>N-Acetylneuraminic acid (Neu5Ac) which is the predominant sialic acid in mammalian cells, is the important target for a pathologic bacteria. And neuraminidase (sialidase) is a kind of glycoside hydrolases, which catalyze the hydrolysis of a glycosidic bond of neuraminic acid, plays important roles in the cell functions such as differentiation, growth, apoptosis and migration, and in the virus replication cycle. A viral neuraminidase is also important for a drug target. So it is important to visualize the location of a neuraminidase activity. For this purpose, some neuraminidase substrates are commercially available and are also useful, but they are not able to stain the location of the neuraminidase activity on a tissue. Therefore we designed the novel neuraminidase substrate which is not fluorescent before enzymatic hydrolysis and is strong fluorescent after yielding water-insoluble BTP which is immobile from the neuraminidase activity position. BTP emits fluorescence through the process called excited state intramolecular proton transfer (ESIPT), so the phenolic proton of BTP is essential for the luminescence. A kind of protective moiety, such as glucose moiety, on phenolic oxygen inhibits the ESIPT fluoresce process. An enzymatic removal of “the protective group” results in the yielding of a strong fluorescent BTP particle and the yielded BTP stains the location of enzyme. We have developed a new artificial substrate for sialidase, which enabled to detect influenza infected cells or location of human colon cancer. This article describes the synthesis and demonstration of histochemical fluorescent staining of influenza infected living cells by detection of sialidase activity.</p>

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