The estimation of dihydropteridine reductase in human blood cells.
The estimation of dihydropteridine reductase in human blood cells.
复制标题
人血细胞中二氢蝶啶还原酶的估计。
DOI：
10.1016/0009-8981(88)90249-5
复制
发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Whiteley,JM
中科院分区：
文献类型：
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作者：
Webber,S;Hural,JA;Whiteley,JM
通过针对大鼠肝酶产生的多克隆抗体在蛋白质印迹中检测人类蛋白质的能力，已经确定了大鼠和人类来源的二氢蝶啶还原酶 (DHPR) 之间的显着同源性。该抗体还与牛、狗和猪肾提取物中的单一蛋白质发生反应，但在兔子中仅检测到微量反应性。通过软激光光密度测定法进行的蛋白质印迹定量表明，响应与纯大鼠肝酶样品以及大鼠和人肝脏粗提物分析中存在的总蛋白成正比。通过该方法分析人血细胞的 DHPR 含量，并将结果与酶测定中测定的水平进行比较。血小板和淋巴细胞的提取物在这两种方法之间显示出良好的相关性，然而，粒细胞表现出高表观酶活性，但在印迹中未检测到 DHPR 蛋白。红细胞提取物显示 DHPR 蛋白水平比活性测量预测的低约 50%。这些结果与通过全血样本酶法测定检测人类 DHPR 缺陷的准确性进行了讨论。
Significant homology between dihydropteridine reductase (DHPR) from rat and human sources has been established by the ability of polyclonal antibodies raised to the rat-liver enzyme to detect the human protein in Western blots. The antibody also reacted with a single protein in bovine, dog and porcine kidney extracts, however, only trace reactivity was detected in rabbit. Quantitation of Western blots by soft laser densitometry showed that the response was proportional to total protein present in analyses of both pure rat-liver enzyme samples and crude extracts of rat and human liver. The DHPR contents of human blood cells were analysed by this method and the results compared to levels determined in enzymatic assays. Extracts of platelets and lymphocytes showed good correlation between these two methods, however, granulocytes exhibited high apparent enzyme activity but no DHPR protein detectable in blots. Erythrocyte extracts showed approximately 50% lower DHPR protein levels than predicted by activity measurements. These results are discussed in relation to the accuracy of detecting DHPR deficiencies in humans by enzymatic assay of whole blood samples.