Ganesh Subramaniam, Tsurutani Naomi, Suzuki Toshimitsu, Hoshii Yoshinobu, Ishihara Tokuhiro, Delgado-Escueta Antonio V, Yamakawa Kazuhiro
Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
Biochem Biophys Res Commun. 2004 Jan 23;313(4):1101-9. doi: 10.1016/j.bbrc.2003.12.043.
Lafora's disease (LD) is an autosomal recessive and fatal form of epilepsy with onset in late childhood or adolescence. One of the characteristic features of LD pathology is the presence of periodic acid-Schiff (PAS) positive Lafora inclusion bodies. Lafora bodies are present primarily in neurons, but they have also been found in other organs. Histochemical and biochemical studies have indicated that Lafora bodies are composed mainly of polysaccharides. The LD gene, EPM2A, encodes a 331 amino acid long protein named laforin that contains an N-terminal carbohydrate-binding domain (CBD) and a C-terminal dual-specificity phosphatase domain (DSPD). Here we demonstrate that the CBD of laforin targets the protein to Lafora inclusion bodies and this property could be evolutionarily conserved. We also tested in vitro the effects of five LD missense mutations on laforin's affinity to Lafora body. While the missense mutant W32G failed to bind to purified Lafora body, four other mutants (S25P, E28L, F88L, and R108C) did not show any effect on the binding affinity. Based on these observations we propose the existence of a laforin-mediated glycogen metabolic pathway regulating the disposal of pathogenic polyglucosan inclusions. This is the first report demonstrating a direct association between the LD gene product and the disease-defining storage product, the Lafora bodies.
拉福拉病(LD)是一种常染色体隐性遗传的致命性癫痫,发病于儿童晚期或青春期。LD病理学的特征之一是存在过碘酸希夫(PAS)阳性的拉福拉包涵体。拉福拉小体主要存在于神经元中,但也在其他器官中被发现。组织化学和生物化学研究表明,拉福拉小体主要由多糖组成。LD基因EPM2A编码一种名为拉福林的331个氨基酸长的蛋白质,该蛋白质含有一个N端碳水化合物结合结构域(CBD)和一个C端双特异性磷酸酶结构域(DSPD)。在此,我们证明拉福林的CBD将该蛋白质靶向拉福拉包涵体,且这一特性可能在进化上是保守的。我们还在体外测试了五个LD错义突变对拉福林与拉福拉小体亲和力的影响。虽然错义突变体W32G未能与纯化的拉福拉小体结合,但其他四个突变体(S25P、E28L、F88L和R108C)对结合亲和力没有任何影响。基于这些观察结果,我们提出存在一种由拉福林介导的糖原代谢途径,该途径调节致病性多聚葡萄糖包涵体的处理。这是首次报道证明LD基因产物与疾病定义性储存产物拉福拉小体之间存在直接关联。