Rapid disappearance of deoxyribose-1-phosphate in platelet derived endothelial cell growth factor/thymidine phosphorylase overexpressing cells
Rapid disappearance of deoxyribose-1-phosphate in platelet derived endothelial cell growth factor/thymidine phosphorylase overexpressing cells
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DOI：
10.1016/s0006-291x(03)00022-6
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发表时间：
2003-02-14
影响因子：
3.1
通讯作者：
Peters, GJ
中科院分区：
文献类型：
--
作者：
de Bruin, M;Smid, K;Peters, GJ
血小板源性内皮细胞生长因子/胸苷磷酸化酶 (PD-ECGF/TP) 催化胸苷 (TdR) 磷酸解为胸腺嘧啶和脱氧核糖 1-磷酸 (dR-1-P)，并具有促血管生成作用，dR-1-P 可能是其中的原因。使用基于嘌呤核苷磷酸化酶的测定发现，TdR 孵育不会增加结肠癌细胞系 Colo320 及其 PD-ECGF/TP 转染变体 Colo320TP1 中 dR-1-P 的积累。该测定在 25,000 pmol dR-1-P 范围内呈线性，并且从细胞提取物中完全回收 dR-1-P。胸腺嘧啶产量与测得的 dR-1-P 水平存在巨大差异，发现 dR-1-P 预期值的 0.05%，表明 dR-1-P 快速消失。然而，在细胞提取物中，TdR 孵育增加了 dR-1-P（可通过捕获测量），而胸苷磷酸化酶抑制剂可抑制 dR-1-P。直接添加到细胞提取物中的dR-1-P在5-10分钟内消失。总之，Colo320TP1 细胞产生大量 dR-1-P，但很快消失，因此不会导致这些细胞中 dR-1-P 的净积累。 (C) 2003 年爱思唯尔科学（美国）。版权所有。
Platelet derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP) catalyzes the phosphorolysis of thymidine (TdR) to thymine and deoxyribose-1-phosphate (dR-1-P) and has a pro-angiogenic effect for which dR-1-P may be responsible. Using a purine nucleoside phosphorylase based assay it was found that TdR incubation did not increase dR-1-P accumulation in colon cancer cell line Colo320 and its PD-ECGF/TP transfected variant Colo320TP1. The assay was linear up to 25,000 pmol dR-1-P with complete recovery of dR-1-P from cellular extracts. There was a huge discrepancy between thymine production and the measured dR-1-P level, 0.05% of the expected value for dR-1-P was found, indicating that there was a rapid disappearance of dR-1-P. However, in cellular extracts, TdR incubation increased dR-1-P, measurable by trapping, which was inhibited by a thymidine phosphorylase inhibitor. dR-1-P directly added to cellular extracts disappeared within 5-10 min. In conclusion, large amounts of dR-1-P are produced by Colo320TP1 cells, which rapidly disappear thus not resulting in a net accumulation of dR-1-P in these cells. (C) 2003 Elsevier Science (USA). All rights reserved.