人类和牛ATP的识别:棒子(I)就adenosyltransferase互补基于互补的细菌突变体。
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公园SD, Leal NA Kima PE、Bobik助教
人类和牛ATP的识别:棒子(I)就adenosyltransferase互补基于互补的细菌突变体。
J生物化学杂志。2003年3月14日,278 (11):9227 - 34。Epub 2003年1月3。
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12514191 (在PubMed]
- 文摘
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在人类中,缺陷在辅酶B12-dependent methylmalonyl-CoA变位酶(MCM)导致methylmalonyl酸尿,一种罕见的疾病,在新生儿往往是致命的。这样的缺陷会导致来自天生的MCM结构基因中的错误或突变影响膳食维生素B12的同化到辅酶B12 (Ado-B12),对MCM所需的辅助因子。ATP:棒子(I)就adenosyltransferase (ATR)催化氰钴胺素转化为Ado-B12终端一步。大量证据表明,这种酶遗传缺陷导致methylmalonyl酸尿,但是相应的ATR基因没有被确认。在这里,我们报告的鉴定牛和人类ATR的互补以及相应的人类基因。一个牛肝cDNA表达文库筛选克隆,补充一个ATR-deficient菌株对颜色形成醛指标中,和四个阳性克隆被孤立。两个克隆的DNA序列测定,发现是相同的。序列相似性搜索被用来确定一个人类同源互补脱氧核糖核酸(89%身份)和其相应的基因位于染色体十二世。牛和人类的互补独立克隆和表达的大肠杆菌。酶化验表明,表达菌株生产87和98 nmol /分钟/毫克ATR活动,分别。 These specific activities are in line with values reported previously for bacterial ATR enzymes. Subsequent studies showed that the human cDNA clone complemented an ATR-deficient bacterial mutant for Ado-B12-dependent growth on 1,2-propanediol. This demonstrated that the human ATR is active under physiological conditions albeit in a heterologous host. In addition, Western blots were used to show that ATR expression is altered in cell lines derived from cblB methylmalonyl aciduria patients compared with cell lines from normal individuals. We propose that inborn errors in the human ATR gene identified here result in methylmalonyl aciduria. The identification of genes involved in this disorder will allow improvements in the diagnosis and treatment of this serious disease.