MOLECULAR-BASIS OF CHRONIC NON-SPHEROCYTIC HEMOLYTIC-ANEMIA - A NEW G6PD VARIANT (393 ARG-]HIS) WITH ABNORMAL KMG6P AND MARKED INVIVO INSTABILITY
MOLECULAR-BASIS OF CHRONIC NON-SPHEROCYTIC HEMOLYTIC-ANEMIA - A NEW G6PD VARIANT (393 ARG-]HIS) WITH ABNORMAL KMG6P AND MARKED INVIVO INSTABILITY
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DOI：
10.1111/j.1365-2141.1992.tb06409.x
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发表时间：
1992-01-01
影响因子：
6.5
通讯作者：
BATTISTUZZI, G
中科院分区：
文献类型：
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作者：
FILOSA, S;CALABRO, V;BATTISTUZZI, G
葡萄糖-6-磷酸脱氢酶（G6 PD）的80多种遗传变异与慢性非球形红细胞溶血性贫血（CNSHA）有关。为了帮助澄清这种关联的分子基础，我们对受CNSHA影响的两个G6 PD缺陷兄弟进行了详细的生化和遗传表征。来自这两个患者的G6 PD具有改变的电泳迁移率、异常升高的G6 P米氏常数（K（m））以及在体内和体外的极端不稳定性。通过与已发表的信息进行比较，我们发现这是一种新的G6 PD变体，我们将其命名为G6 PD Porcine。该基因的整个编码区已被测序，在外显子X的1178位发现了一个单点突变，即G->A转换，导致多肽链中第393位残基的精氨酸被组氨酸取代。通过聚合酶链反应（PCR）扩增，然后进行诊断性限制性内切酶分析和等位基因特异性寡核苷酸杂交，我们已经证明了该突变在患者家族中的遗传。我们的研究结果支持G6 PD基因突变与CNSHA之间存在因果关系的观点。我们的数据，结合文献中以前的数据，表明G6 PD的三维结构是这样的，以引起其两个底物，G6 P和NADP的结合的相互作用。
More than 80 genetic variants of glucose-6-phosphate dehydrogenase (G6PD) are associated with chronic non-spherocytic haemolytic anaemia (CNSHA). In order to help clarify the molecular basis of this association, we have carried out a detailed biochemical and genetic characterization of two G6PD deficient brothers affected by CNSHA. The G6PD from the two patients has altered electrophoretic mobility, abnormally elevated Michaelis constant (K(m)) for G6P, and extreme instability in vivo and in vitro. By comparison with published information we found that this is a new G6PD variant which we have designated G6PD Portici. The entire coding region of the gene has been sequenced, and a single point mutation, a G-->A transition, was found at position 1178 in exon X, causing a substitution of histidine for arginine at residue 393 in the polypeptide chain. By polymerase chain reaction (PCR) amplification followed by diagnostic restriction enzyme analysis and allele-specific oligonucleotide hybridization we have demonstrated the inheritance of this mutation in the patient's family. Our results support the notion of a causative link between this mutation in the G6PD gene and CNSHA. Our data, in combination with previous data in the literature, suggest that the three-dimensional structure of G6PD is such as to cause interaction in the binding of its two substrates, G6P and NADP.