Stimuli-responsive "activatable" reactive tags are applicable to selective labeling of biomolecules in a defined area or environment in living systems, yielding new insights into cellular processes through molecular imaging and fishing. Here, we developed a chemically activatable alkyne tag that can be incorporated into biological molecules and labeled with azide-tagged functional molecules through the alkyne-azide cycloaddition "click" reaction after chemical activation. Formation of the alkyne tag from the precursor moiety was confirmed to proceed in physiological aqueous media and was particularly enhanced under mildly acidic pH. The tag was successfully applied to low-pH sensitive labeling of a cholesterol analogue with azide-tagged biotin on living mammalian cells. Our results provided proof of principle that the present activatable alkyne tag can be used as a tool to selectively analyze molecules of interest in low-pH regions in living systems.
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