Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge CB2 0XY, United Kingdom.
Proc Natl Acad Sci U S A. 2011 Sep 13;108(37):15169-73. doi: 10.1073/pnas.1105639108. Epub 2011 Aug 29.
Krabbe disease is a devastating neurodegenerative disease characterized by widespread demyelination that is caused by defects in the enzyme galactocerebrosidase (GALC). Disease-causing mutations have been identified throughout the GALC gene. However, a molecular understanding of the effect of these mutations has been hampered by the lack of structural data for this enzyme. Here we present the crystal structures of GALC and the GALC-product complex, revealing a novel domain architecture with a previously uncharacterized lectin domain not observed in other hydrolases. All three domains of GALC contribute residues to the substrate-binding pocket, and disease-causing mutations are widely distributed throughout the protein. Our structures provide an essential insight into the diverse effects of pathogenic mutations on GALC function in human Krabbe variants and a compelling explanation for the severity of many mutations associated with fatal infantile disease. The localization of disease-associated mutations in the structure of GALC will facilitate identification of those patients that would be responsive to pharmacological chaperone therapies. Furthermore, our structure provides the atomic framework for the design of such drugs.
克拉伯病是一种破坏性的神经退行性疾病,其特征是广泛的脱髓鞘,由半乳糖脑苷脂酶(GALC)的酶缺陷引起。在 GALC 基因中已经发现了致病突变。然而,由于缺乏这种酶的结构数据,对这些突变的分子影响的理解一直受到阻碍。在这里,我们展示了 GALC 的晶体结构和 GALC-产物复合物的结构,揭示了一种新颖的结构域架构,其中包含一个以前未被表征的凝集素结构域,在其他水解酶中未观察到。GALC 的所有三个结构域都对底物结合口袋有贡献,致病突变广泛分布于整个蛋白质中。我们的结构为人类克拉伯病变体中致病性突变对 GALC 功能的多种影响提供了重要的见解,并为与致命婴儿疾病相关的许多突变的严重程度提供了令人信服的解释。GALC 结构中疾病相关突变的定位将有助于识别对药理伴侣治疗有反应的患者。此外,我们的结构为这类药物的设计提供了原子框架。