Identification of Tissue-Restricted Bioreaction Suitable for in Vivo Targeting by Fluorescent Substrate Library-Based Enzyme Discovery

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• 作者：Kentaro Yoshioka ; Toru Komatsu ; Akihiro Nakada ; Jun Onagi ; Yugo Kuriki ; Mitsuyasu Kawaguchi ; Takuya Terai ; Tasuku Ueno ; Kenjiro Hanaoka ; Tetsuo Nagano ; Yasuteru Urano
• 刊名：Journal of the American Chemical Society
• 出版年：2015
• 出版时间：September 30, 2015
• 年：2015
• 卷：137
• 期：38
• 页码：12187-12190
• 全文大小：302K
• ISSN：1520-5126

文摘

Tissue-restricted bioreactions can be utilized to design chemical-biological tools and prodrugs. We have developed a fluorescent-substrate-library-based enzyme discovery approach to screen tissue extracts for enzymatic activities of interest. Assay-positive candidate proteins were identified by diced electrophoresis gel assay followed by peptide mass fingerprinting. We discovered that pyruvyl anilide is specifically hydrolyzed by carboxylesterase 2 (CES2), which is predominantly localized in the liver and kidney. We show that the pyruvyl targeting group/CES2 enzyme pair can be used to deliver the 7-amino-4-methylcoumarin fluorophore specifically to the liver and kidney in vivo. Our screening approach should be useful to find other masking group/enzyme pairs suitable for development of fluorescent substrates and prodrugs.