Scher M G, Burton W A, Waechter C J
J Neurochem. 1980 Oct;35(4):844-9. doi: 10.1111/j.1471-4159.1980.tb07081.x.
When calf brain membrane preparations containing endogenous dolichyl [32P]monophosphate (Dol-32P), prelabeled enzymatically by [gamma-32P]-CTP, are incubated with unlabeled UDP-glucose, the formation of a mild acid-labile [32P]phosphoglucolipid is observed. The biosynthesis of the [32P]phosphoglucolipid is dependent on the concentration of UDP-glucose added, and no [32P]phosphoglycolipid appeared when UDP-glucose was replaced by ADP-glucose, UDP-xylose, UDP-galactose, UDP-mannose, or UDP-glucuronic acid. The 32P-labeled product formed by the UDP-glucose-dependent reaction is chemically and chromatographically identical to glucosylphosphoryldolichol. Several enzymatic parameters of the glucosylation of the specific pool of Dol-P, synthesized by the CTP-mediated kinase, and the total available pool of Dol-P have been compared by a double-label assay utilizing endogenous, prelabeled Dol-32P and UDP-[3H]glucose as substrates.
当含有通过[γ-32P]-CTP酶促预标记的内源性多萜醇[32P]单磷酸酯（Dol-32P）的小牛脑膜制剂与未标记的UDP-葡萄糖一起温育时，会观察到一种对弱酸不稳定的[32P]磷酸葡萄糖脂的形成。[32P]磷酸葡萄糖脂的生物合成取决于所添加的UDP-葡萄糖的浓度，当UDP-葡萄糖被ADP-葡萄糖、UDP-木糖、UDP-半乳糖、UDP-甘露糖或UDP-葡萄糖醛酸取代时，未出现[32P]磷酸糖脂。由UDP-葡萄糖依赖性反应形成的32P标记产物在化学和色谱上与葡萄糖基磷酸多萜醇相同。利用内源性预标记的Dol-32P和UDP-[3H]葡萄糖作为底物，通过双标记测定法比较了由CTP介导的激酶合成的Dol-P特定池和Dol-P总可用池的糖基化的几个酶学参数。
