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Metabolic labeling strategy for cellular biosynthesis of O-GlcNDAz-modified proteins. Cellular GlcNAc can be converted to UDP-GlcNAc via three enzymatic steps catalyzed by N-acetylglucosamine kinase (NAGK), N-acetylglucosamine-phosphate mutase (AGM1), and UDP-GlcNAc pyrophosphorylase (AGX1 or AGX2). HeLa cells expressing AGX1(F383G) and cultured with Ac3GlcNDAz-1-P(Ac-SATE)2 efficiently produced UDP-GlcNDAz.
