The pharmacological chaperone isofagomine increases the activity of the Gaucher disease L444P mutant form of beta-glucosidase.
The pharmacological chaperone isofagomine increases the activity of the Gaucher disease L444P mutant form of beta-glucosidase.
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DOI：
10.1111/j.1742-4658.2010.07588.x
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发表时间：
2010-04
期刊：
影响因子：
--
通讯作者：
Valenzano KJ
中科院分区：
文献类型：
--
作者：
Khanna R;Benjamin ER;Pellegrino L;Schilling A;Rigat BA;Soska R;Nafar H;Ranes BE;Feng J;Lun Y;Powe AC;Palling DJ;Wustman BA;Schiffmann R;Mahuran DJ;Lockhart DJ;Valenzano KJ
Gaucher病是由编码溶酶体酶β-葡萄糖苷酶（GCASE）的基因中的突变引起的，我们先前已经表明，小分子药物链酮同产量（IFG）结合了N370S，导致了N370S的GCASE。 。在细胞裂解物中，使用糖蛋白含量，gcase-免疫接种或通过在分析前在无IFG的培养基中孵育过夜，增加了成纤维细胞的作用，增加了大约2倍。酶体孵化也增加了溶酶体的运输和完整细胞中L444p GCASE的原位活性，这是通过降低内源性葡萄糖基酶水平来衡量的。 IFG清除对表达鼠L444p gcase的小鼠的溶酶体GCASE活性的重要性，IFG的口服导致疾病与疾病相关的组织中的GCASE活性显着增加（2-5倍）。 IFG给药显着降低了血浆壳蛋白III和IgG水平，24周的给药可显着降低脾脏和肝脏的重量，这些数据表明IFG可以增加细胞和组织中L444P GCASE的溶酶体活性。可用并分布到包括大脑在内的多个时机中，因此可能对神经性和非神经性神经疾病的患者进行治疗评估。
Gaucher disease is caused by mutations in the gene that encodes the lysosomal enzyme acid β-glucosidase (GCase). We have shown previously that the small molecule pharmacological chaperone isofagomine (IFG) binds and stabilizes N370S GCase, resulting in increased lysosomal trafficking and cellular activity. In this study, we investigated the effect of IFG on L444P GCase. Incubation of Gaucher patient-derived lymphoblastoid cell lines (LCLs) or fibroblasts with IFG led to approximately 3.5- and 1.3-fold increases in L444P GCase activity, respectively, as measured in cell lysates. The effect in fibroblasts was increased approximately 2-fold using glycoprotein-enrichment, GCase-immunocapture, or by incubating cells overnight in IFG-free media prior to assay, methods designed to maximize GCase activity by reducing IFG carryover and inhibition in the enzymatic assay. IFG incubation also increased the lysosomal trafficking and in situ activity of L444P GCase in intact cells, as measured by reduction in endogenous glucosylceramide levels. Importantly, this reduction was seen only following three-day incubation in IFG-free media, underscoring the importance of IFG removal to restore lysosomal GCase activity. In mice expressing murine L444P GCase, oral administration of IFG resulted in significant increases (2- to 5-fold) in GCase activity in disease-relevant tissues, including brain. Additionally, eight-week IFG administration significantly lowered plasma chitin III and IgG levels, and 24-week administration significantly reduced spleen and liver weights. Taken together, these data suggest that IFG can increase the lysosomal activity of L444P GCase in cells and tissues. Moreover, IFG is orally available and distributes into multiple tissues, including brain, and may thus merit therapeutic evaluation for patients with neuronopathic and non-neuronopathic Gaucher disease.
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影响因子：
158.5
作者：
BARTON, NW;BRADY, RO;YU, KT
通讯作者：
YU, KT
DOI：
10.1089/gte.1997.1.5
发表时间：
1997-06-01
期刊：
GENETIC TESTING
影响因子：
--
作者：
Grabowski, GA
通讯作者：
Grabowski, GA
影响因子：
168.9
作者：
Cox, T;Lachmann, R;Zimran, A
通讯作者：
Zimran, A
影响因子：
14.8
作者：
Lieberman, Raquel L.;Wustman, Brandon A.;Petsko, Gregory A.
通讯作者：
Petsko, Gregory A.
影响因子：
7.7
作者：
Dvir, H;Harel, M;Sussman, JL
通讯作者：
Sussman, JL