Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.
Blood Cells Mol Dis. 2011 Jan 15;46(1):19-26. doi: 10.1016/j.bcmd.2010.07.009. Epub 2010 Aug 21.
GBA3, also known as cytosolic β-glucosidase, is thought to hydrolyze xenobiotic glycosides in man. Deficiency of glucocerebrosidase (GBA), a β-glucosidase degrading glucosylceramide, underlies Gaucher disease. We examined GBA3, which recently was proposed to degrade glucosylceramide and influence the clinical manifestation of Gaucher disease. Recombinant GBA3 was found to hydrolyze artificial substrates such as 4-methylumbelliferyl-β-D-glucoside and C6-NBD-glucosylceramide, but hydrolysis of naturally occurring lipids like glucosylceramide and glucosylsphingosine was hardly detected. Consistent with this, inhibition of GBA3 in cultured cells using a novel inhibitor (alpha-1-C-nonyl-DIX) did not result in an additional increase in glucosylceramide as compared to GBA inhibition alone. Examination of the GBA3 gene led to the identification of a common substitution in its open reading frame (1368T→A), resulting in a truncated GBA3 protein missing the last α-helix of its (β/α)(8) barrel. Both recombinant 1368A GBA3 and 1368A enzyme from spleen of a homozygous individual were found to be inactive. Amongst non-neuronopathic (type 1) Gaucher disease patients, we subsequently identified individuals being wild-type, heterozygous, or homozygous for the GBA3 1368T→A mutation. No correlation was observed between GBA3 1368A/T haplotypes and severity of type 1 Gaucher disease manifestation. In conclusion, GBA3 does not seem to modify type 1 Gaucher disease manifestation.
GBA3,也称为胞质β-葡萄糖苷酶,据认为它能水解人体中的异生物质糖苷。葡萄糖脑苷脂酶(GBA)的缺乏是导致戈谢病的原因,GBA 是一种能降解葡萄糖脑苷脂的β-葡萄糖苷酶。我们研究了 GBA3,它最近被提出能降解葡萄糖脑苷脂并影响戈谢病的临床表现。研究发现重组 GBA3 能水解人工底物,如 4-甲基伞形酮-β-D-葡糖苷和 C6-NBD-葡萄糖脑苷脂,但对天然脂质,如葡萄糖脑苷脂和葡萄糖鞘氨醇的水解作用几乎检测不到。与此一致的是,使用新型抑制剂(α-1-C-壬基-DIX)在培养的细胞中抑制 GBA3,与单独抑制 GBA 相比,不会导致葡萄糖脑苷脂的进一步增加。对 GBA3 基因的检查导致了其开放阅读框(1368T→A)中的一个常见取代的鉴定,导致缺失其(β/α)(8)桶最后一个α-螺旋的截短 GBA3 蛋白。研究发现重组 1368A GBA3 和 1368A 同工酶均没有活性。在非神经病变型(1 型)戈谢病患者中,我们随后鉴定出 GBA3 1368T→A 突变的野生型、杂合子或纯合子个体。在 1 型戈谢病患者中,未观察到 GBA3 1368A/T 单倍型与疾病严重程度之间存在相关性。总之,GBA3 似乎不会改变 1 型戈谢病的临床表现。