Isolation of a hamster cDNA clone coding for a function involved in methotrexate uptake.
Isolation of a hamster cDNA clone coding for a function involved in methotrexate uptake.
复制标题
编码与甲氨蝶呤吸收相关的功能的仓鼠 cDNA 克隆的分离。
DOI：
10.1016/s0021-9258(17)37534-8
复制
发表时间：
1994
期刊：
影响因子：
--
通讯作者：
W. Flintoff
中科院分区：
文献类型：
--
作者：
F. Williams;R. Murray;T. Underhill;W. Flintoff
从中国仓鼠卵巢细胞cDNA表达文库中分离出一个克隆，该克隆补充了叶酸类似物甲氨蝶呤摄取缺陷的突变细胞。当用该克隆转染时，突变细胞重新获得结合和转运药物的能力，因此对其细胞毒性作用变得敏感。该克隆长2314个碱基对，具有1557个碱基对的开放阅读框，编码58 kDa的推定蛋白质。这种新的推定蛋白质具有高含量的疏水残基，并具有其预测的β-折叠形式的二级结构的很大一部分。在野生型细胞系和一个突变细胞系中，该克隆检测到2.5个酶的mRNA，而在另一个突变细胞系中，该信息不存在。数据与编码还原叶酸转运蛋白或与该转运蛋白相互作用的辅助功能的该克隆一致。这是第一次报告的cDNA编码的功能，而不是叶酸结合蛋白，这是参与甲氨蝶呤的运输。
A clone has been isolated from a Chinese hamster ovary cell cDNA expression library that complements mutant cells defective in the uptake of the folate analogue methotrexate. When transfected with this clone, the mutant cells regain the ability to bind and transport the drug and, as a consequence, become sensitive to its cytotoxic action. The clone is 2314 base pairs long and has an open reading frame of 1557 base pairs that codes for a putative protein of 58 kDa. This novel putative protein has a high content of hydrophobic residues and has a large part of its predicted secondary structure in the form of beta-sheets. In the wild-type cell line and in one of the mutant cell lines, this clone detects an mRNA of 2.5 kilobases, while in another mutant cell line, this message is absent. The data are consistent with this clone encoding either the reduced folate transporter or an auxiliary function that interacts with this transporter. This is the first report of a cDNA coding for a function, other than folate-binding protein, that is involved in the transport of methotrexate.