Beutler E, Kuhl W, Gelbart T, Forman L
Scripps Clinic and Research Foundation, Research Institute of Scripps Clinic, La Jolla, California 92037.
J Biol Chem. 1991 Mar 5;266(7):4145-50.
Over 400 supposedly biochemically and genetically distinct variants of glucose-6-phosphate dehydrogenase (G6PD) have been described in the past. In order to investigate these variants at the DNA sequence level we have now determined the relevant sequences of introns of G6PD and describe a method which allows us to rapidly determine the sequence of the entire coding region of G6PD. This technique was applied to six variants that cause G6PD deficiency to be functionally so severe as to result in nonspherocytic hemolytic anemia. Although the patients were all unrelated, G6PD Marion, Gastonia, and Minnesota each had identical mutations, a G----T at nucleotide (nt) 637 in exon 6 leading to a Val----Leu substitution at amino acid 213. The mutations of Nashville and Anaheim were identical to each other, viz. G----A at nt 1178 in exon 10 producing a Arg----His substitution at amino acid 393. G6PD Loma Linda had a C----A substitution at nt 1089 in exon 10, producing a Asn----Lys change at amino acid 363. The results confirm our earlier results suggesting that the NADP-binding site is in a small region of exon 10 and suggest the possibility that this area is also concerned with the binding of glucose-6-P.
过去已描述了400多种葡萄糖-6-磷酸脱氢酶（G6PD）在生化和遗传方面据称不同的变体。为了在DNA序列水平上研究这些变体，我们现在已确定了G6PD内含子的相关序列，并描述了一种能让我们快速确定G6PD整个编码区序列的方法。该技术应用于六种导致G6PD缺乏症在功能上严重到足以引发非球形红细胞溶血性贫血的变体。尽管这些患者毫无亲缘关系，但G6PD Marion、Gastonia和Minnesota每种变体都有相同的突变，即外显子6中核苷酸（nt）637处的G→T，导致第213位氨基酸由缬氨酸变为亮氨酸。Nashville和Anaheim的突变彼此相同，即外显子10中nt 1178处的G→A，导致第393位氨基酸由精氨酸变为组氨酸。G6PD Loma Linda在外显子10的nt 1089处有一个C→A替换，导致第363位氨基酸由天冬酰胺变为赖氨酸。这些结果证实了我们早期的结果，表明NADP结合位点在外显子10的一个小区域内，并提示该区域也可能与葡萄糖-6-P的结合有关。
