Diels鈥揂lder Cycloaddition for Fluorophore Targeting to Specific Proteins inside Living Cells

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• 作者：Daniel S. Liu ; Anupong Tangpeerachaikul ; Ramajeyam Selvaraj ; Michael T. Taylor ; Joseph M. Fox ; Alice Y. Ting
• 刊名：The Journal of the American Chemical Society
• 出版年：2012
• 出版时间：January 18, 2012
• 年：2012
• 卷：134
• 期：2
• 页码：792-795
• 全文大小：269K
• 年卷期：v.134,no.2(January 18, 2012)
• ISSN：1520-5126

文摘

The inverse-electron-demand Diels鈥揂lder cycloaddition between trans-cyclooctenes and tetrazines is biocompatible and exceptionally fast. We utilized this chemistry for site-specific fluorescence labeling of proteins on the cell surface and inside living mammalian cells by a two-step protocol. Escherichia coli lipoic acid ligase site-specifically ligates a trans-cyclooctene derivative onto a protein of interest in the first step, followed by chemoselective derivatization with a tetrazine鈥揻luorophore conjugate in the second step. On the cell surface, this labeling was fluorogenic and highly sensitive. Inside the cell, we achieved specific labeling of cytoskeletal proteins with green and red fluorophores. By incorporating the Diels鈥揂lder cycloaddition, we have broadened the panel of fluorophores that can be targeted by lipoic acid ligase.