Taljanidisz J, Sasvari-Szekely M, Spasokutotskaja T, Antoni F, Staub M
Biochim Biophys Acta. 1986 Mar 14;885(3):266-71. doi: 10.1016/0167-4889(86)90241-7.
The total uptake, phosphorylation and incorporation of thymidine (dThd) and deoxycytidine (dCyd) were compared in intact and reversibly permeabilized human tonsillar lymphocytes. The total uptake of [3H]dThd was lower than that of [5-3H]dCyd, but almost all of [3H]dThd was incorporated into DNA. However, the main part of [5-3H]dCyd taken up by the lymphocytes was found in the pool as phosphorylated nucleoside (55%), and only a smaller part (13%) was incorporated into DNA. Phosphorylated nucleosides were determined by DEAE-cellulose sheets in the ethanol-soluble fraction of the cells. The reversible permeabilization of lymphocytes by Dextran T-150 destroys totally the [3H]dThd incorporation, while [5-3H]dCyd incorporation decreased only to 60% of intact cells. During permeabilization the phosphorylation of both nucleosides increased severalfold. After permeabilization all [3H]dThd was in dTMP form, while [5-3H]dCyd was also found in dCDP (3%) and dCTP (38%) form. In the meanwhile, 22% of thymidine kinase, 63% of deoxycytidine kinase and 98% of DNA polymerase activity were measured in permeabilized cells as compared to intact cells. The results suggest different relationships between the lymphocyte plasma membrane and the salvage pathways of the two pyrimidine nucleosides.
在完整的和可逆通透的人扁桃体淋巴细胞中，比较了胸苷（dThd）和脱氧胞苷（dCyd）的总摄取、磷酸化及掺入情况。[3H]dThd的总摄取量低于[5-3H]dCyd，但几乎所有的[3H]dThd都掺入了DNA。然而，淋巴细胞摄取的[5-3H]dCyd的主要部分存在于磷酸化核苷池中（55%），只有较小一部分（13%）掺入了DNA。通过DEAE-纤维素薄板在细胞的乙醇可溶部分中测定磷酸化核苷。用葡聚糖T-150对淋巴细胞进行可逆通透处理完全破坏了[3H]dThd的掺入，而[5-3H]dCyd的掺入仅降至完整细胞的60%。在通透处理过程中，两种核苷的磷酸化均增加了几倍。通透处理后，所有的[3H]dThd都呈dTMP形式，而[5-3H]dCyd也以dCDP（3%）和dCTP（38%）形式存在。与此同时，与完整细胞相比，在通透处理的细胞中检测到22%的胸苷激酶、63%的脱氧胞苷激酶和98%的DNA聚合酶活性。结果表明淋巴细胞质膜与两种嘧啶核苷的补救途径之间存在不同的关系。
