Pharmacological Chaperones for the Treatment of α-Mannosidosis
Pharmacological Chaperones for the Treatment of α-Mannosidosis
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DOI：
10.1021/acs.jmedchem.9b00153
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发表时间：
2019-06-27
影响因子：
7.3
通讯作者：
Ortiz Mellet, Carmen
中科院分区：
文献类型：
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作者：
Risquez-Cuadro, Rocio;Matsumoto, Reimi;Ortiz Mellet, Carmen
α-甘露糖苷沉积症（AM）由溶酶体α-甘露糖苷酶（LAMAN）活性缺陷和随后的底物在溶酶体中积累引起，导致严重的病理学。许多AM致病突变损害酶折叠，可以用专门设计的药理学伴侣（PC）来拯救。我们发现，PC结合的LAMAN糖基结合基序的基础上的5 N，6 O-oxomethylidenemanojirimycin（OMJ）糖模拟核心和不同的糖苷配基，在单价或多价显示，引发的结合模式，涉及糖苷配基和非糖苷配基酶区域，加强蛋白质的折叠和稳定的潜力。多价衍生物表现出有效的酶抑制，一般占上风的伴侣效应。相反，具有LAMAN糖苷配基结合面积拟合取代基的单价OMJ衍生物证明在AM患者成纤维细胞和/或转染的MAN 2B 1-KO细胞中作为几种突变LAMAN形式的活性增强剂是有效的。这转化为内体/溶酶体功能的显著改善，不仅恢复了主要的LAMAN底物积累，而且还恢复了额外的下游后果，如胆固醇积累。
alpha-Mannosidosis (AM) results from deficient lysosomal a-mannosidase (LAMAN) activity and subsequent substrate accumulation in the lysosome, leading to severe pathology. Many of the AM-causative mutations compromise enzyme folding and could be rescued with purpose-designed pharmacological chaperones (PCs). We found that PCs combining a LAMAN glycone-binding motif based on the 5N,6O-oxomethylidenemannojirimycin (OMJ) glycomimetic core and different aglycones, in either mono- or multivalent displays, elicit binding modes involving glycone and non-glycone enzyme regions that reinforce the protein folding and stabilization potential. Multivalent derivatives exhibited potent enzyme inhibition that generally prevailed over the chaperone effect. On the contrary, monovalent OMJ derivatives with LAMAN aglycone binding area-fitting substituents proved effective as activity enhancers for several mutant LAMAN forms in AM patient fibroblasts and/or transfected MAN2B1-KO cells. This translated into a significant improvement in endosomal/lysosomal function, reverting not only the primary LAMAN substrate accumulation but also the additional downstream consequences such as cholesterol accumulation.