Eriksson G, Särnstrand B, Malmström A
Arch Biochem Biophys. 1984 Dec;235(2):692-8. doi: 10.1016/0003-9861(84)90245-5.
Nucleotides and sugar nucleotides were extracted from cultures of human fibroblasts with perchloric acid, separated by isotachophoresis, and quantified by uv absorption analysis at 254 nm. ATP (936 pmol/micrograms DNA) was, as expected, the dominating nucleotide pool. The energy charge was estimated to 0.9. The UDP-N-acetylhexosamine pool was also a very prominent compound (596 pmol/micrograms DNA). After incubation of fibroblasts with [3H]glucosamine, more than 95% of the acid-soluble radioactivity was found in the UDP-N-acetylhexosamine pool. Incubation with [35S]sulfate resulted in the incorporation of [35S]sulfate into 3'-phosphoadenosine-5'-phosphosulfate (PAPS). The latter could, however, only be measured as radioactivity, as the amount was too small to be quantified as total mass. Pulse-labeling of fibroblasts with [35S]sulfate and [3H]glucosamine from 5 min to 16 h showed that [35S]PAPS was equilibrated in less than 10 min, while [3H]glucosamine required a longer time, 2-4 h, to attain a steady state with UDP-N-acetylhexosamine. [14C]Glucose required approximately the same time as [3H]glucosamine to reach steady state with UDP-acetylhexosamine, which suggests that the reason for the long equilibration time is the slow turnover of this pool.
用高氯酸从人成纤维细胞培养物中提取核苷酸和糖核苷酸，通过等速电泳进行分离，并在254nm处通过紫外吸收分析进行定量。如预期的那样，ATP（936皮摩尔/微克DNA）是主要的核苷酸库。能量电荷估计为0.9。UDP-N-乙酰己糖胺库也是一种非常突出的化合物（596皮摩尔/微克DNA）。用[3H]葡萄糖胺孵育人成纤维细胞后，超过95%的酸溶性放射性物质存在于UDP-N-乙酰己糖胺库中。用[35S]硫酸盐孵育导致[35S]硫酸盐掺入3'-磷酸腺苷-5'-磷酸硫酸酯（PAPS）。然而，由于其含量太小，无法作为总质量进行定量，因此只能将后者测量为放射性。用[35S]硫酸盐和[3H]葡萄糖胺对人成纤维细胞进行5分钟至16小时的脉冲标记表明，[35S]PAPS在不到10分钟内达到平衡，而[3H]葡萄糖胺需要更长的时间（2-4小时）才能与UDP-N-乙酰己糖胺达到稳态。[14C]葡萄糖与UDP-乙酰己糖胺达到稳态所需的时间与[3H]葡萄糖胺大致相同，这表明平衡时间长的原因是该库的周转缓慢。
