Insights into post-translational processing of beta-galactosidase in an animal model resembling late infantile human G-gangliosidosis.
Insights into post-translational processing of beta-galactosidase in an animal model resembling late infantile human G-gangliosidosis.
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DOI：
10.1111/j.1582-4934.2007.00204.x
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发表时间：
2008-09
影响因子：
5.3
通讯作者：
Baumgärtner W
中科院分区：
文献类型：
--
作者：
Kreutzer R;Kreutzer M;Pröpsting MJ;Sewell AC;Leeb T;Naim HY;Baumgärtner W
GM 1-神经节苷脂沉积症是一种由β-半乳糖苷酶活性缺乏引起的溶酶体储存疾病。人类GM 1-神经节苷脂沉积症根据临床发病年龄和特定生化指标分为三种形式。在本研究中，犬模型II型晚期婴儿人GM 1-神经节苷脂沉积症进行了详细的研究“在体外”。为了更好地了解GM 1-神经节苷脂沉积症的分子发病机制，本研究着重于分析β-半乳糖苷酶的分子事件和随后的细胞内蛋白运输。在犬模型中，遗传缺陷导致mRNA转录物中外显子15的排除或包含以及两种突变前体蛋白的翻译。研究了这些突变蛋白的细胞内定位、加工和酶活性。所获得的结果表明，由外显子15和16编码的β-半乳糖苷酶C-末端对于正确的C-末端蛋白水解加工和酶活性是必需的，但不影响到溶酶体的正确路由。两种突变蛋白质前体都是酶失活的，但被转运到溶酶体，清楚地表明外显子15和16编码的氨基酸序列对于正确折叠和与保护性蛋白/组织蛋白酶A结合是必需的，而到溶酶体的路线不受影响。因此，研究的犬模型是人类晚期婴儿型的适当动物模型，代表了测试人类和犬GM 1-神经节苷脂沉积症基因治疗方法的通用系统。
GM1-gangliosidosis is a lysosomal storage disorder caused by a deficiency of ß-galactosidase activity. Human GM1-gangliosidosis has been classified into three forms according to the age of clinical onset and specific biochemical parameters. In the present study, a canine model for type II late infantile human GM1-gangliosidosis was investigated ‘in vitro’ in detail. For a better understanding of the molecular pathogenesis underlying GM1-gangliosidosis the study focused on the analysis of the molecular events and subsequent intracellular protein trafficking of β-galactosidase. In the canine model the genetic defect results in exclusion or inclusion of exon 15 in the mRNA transcripts and to translation of two mutant precursor proteins. Intracellular localization, processing and enzymatic activity of these mutant proteins were investigated. The obtained results suggested that the β-galactosidase C-terminus encoded by exons 15 and 16 is necessary for correct C-terminal proteolytic processing and enzyme activity but does not affect the correct routing to the lysosomes. Both mutant protein precursors are enzymatically inactive, but are transported to the lysosomes clearly indicating that the amino acid sequences encoded by exons 15 and 16 are necessary for correct folding and association with protective protein/cathepsin A, whereas the routing to the lysosomes is not influenced. Thus, the investigated canine model is an appropriate animal model for the human late infantile form and represents a versatile system to test gene therapeutic approaches for human and canine GM1-gangliosidosis.
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DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
影响因子：
3.9
作者：
Caciotti, Anna;Donati, Maria Alice;Morrone, Amelia
通讯作者：
Morrone, Amelia
影响因子：
4.1
作者：
HUBERT, JJ;OBRIEN, JS
通讯作者：
OBRIEN, JS
影响因子：
4.2
作者：
Müller, G;Baumgärtner, W;Kustermann-Kuhn, B
通讯作者：
Kustermann-Kuhn, B
影响因子：
2.4
作者：
Müller, G;Alldinger, S;Baumgärtner, W
通讯作者：
Baumgärtner, W