Saha N, Hong S H, Low P S, Tay J S
Department of Paediatrics, Faculty of Medicine, National University of Singapore.
Hum Hered. 1995 Sep-Oct;45(5):253-7. doi: 10.1159/000154308.
Biochemical characteristics were determined for four common molecular variants of glucose-6-phosphate dehydrogenase (G6PD) deficiency and 10 non-deficient Chinese males in Singapore. The variants included one Mediterranean (nt563 C-->T), two Canton (G-->T at nt1376) and one each of Kaiping (1388 G-->A) and Chinese-5 (1024 C-->T) variants. Molecular identification was carried out by amplication of genomic DNA with specific oligonucleotide primers followed by digestion with restriction enzymes that recognize artificially created or naturally occurring restriction sites. All the variants had low enzyme activity in red cells (0.2-0.6 IU/g Hb). All but the Chinese-5 variant (nt1024) had a normal Km for NADP (7-10 microM). The Mediterranean variant had a high utilization of deamino-NADP (296%), followed by the Canton variant 1376 substitution (131%). The Km for glucose-6-phosphate was low in the Mediterranean and 1376 variant (18-40 microM) but high in the 1024 substitution (104 microM). Electrophoretic mobility in TEB buffer (pH 8.6) was slightly faster (103%) for the 1376 mutation while 100% for all the others. All but the 1024 substitution had increased analogue utilization for galactose-6-phosphate and 2-deoxy-glucose-6-phosphate (58 and 68% for the Mediterranean mutation and 14-23% for the 1376 and 1388 substitutions, respectively), and reduced heat stability.
对新加坡4名葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症的常见分子变体患者及10名非缺乏症中国男性进行了生化特征测定。这些变体包括1种地中海型(nt563 C→T)、2种广东型(nt1376处G→T)、1种开平型(1388 G→A)和1种中国-5型(1024 C→T)变体。通过用特异性寡核苷酸引物扩增基因组DNA,然后用识别人工创建或天然存在的限制性位点的限制性酶进行消化来进行分子鉴定。所有变体的红细胞中酶活性均较低(0.2 - 0.6 IU/g Hb)。除中国-5型变体(nt1024)外,所有变体对NADP的Km值均正常(7 - 10 microM)。地中海型变体对脱氨基-NADP的利用率较高(296%),其次是广东型变体1376替代型(131%)。地中海型和1376变体中葡萄糖-6-磷酸的Km值较低(18 - 40 microM),但在1024替代型中较高(104 microM)。在TEB缓冲液(pH 8.6)中,1376突变体的电泳迁移率略快(103%),而其他所有变体均为100%。除1024替代型外,所有变体对6-磷酸半乳糖和2-脱氧-6-磷酸葡萄糖的类似物利用率均增加(地中海型突变分别为58%和68%,1376和1388替代型分别为14 - 23%),且热稳定性降低。