Functional studies of new GLA gene mutations leading to conformational Fabry disease.
Functional studies of new GLA gene mutations leading to conformational Fabry disease.
复制标题
DOI：
10.1016/j.bbadis.2009.11.003
复制
发表时间：
2010-02
影响因子：
6.2
通讯作者：
Morrone, A.
中科院分区：
文献类型：
--
作者：
Filoni, C.;Caciotti, A.;Carraresi, L.;Cavicchi, C.;Parini, R.;Antuzzi, D.;Zampetti, A.;Feriozzi, S.;Poisetti, P.;Garman, S. C.;Guerrini, R.;Zammarchi, E.;Donati, M. A.;Morrone, A.
法布里病(FD)是一种X连锁的多系统溶酶体疾病，由α-半乳糖苷酶基因突变引起。到目前为止，在已确定的450个遗传损伤中，只有几个被体外表达研究所表征。因此，在FD患者中新发现的导致α-半乳糖苷酶(GAL-A)氨基酸替换或内含子改变的GLA核苷酸变异的意义可能不确定。我们在两名男性和一名女性中发现了三个GLA突变：C155G和Gt；A(p.C52Y)、C548G和Gt；C(p.G183A)、C647A和Gt；G(p.Y216C)，并进行了体外表达研究和Western Blot分析，以阐明它们的功能效应。在所有过表达的突变系统中，Gal-A活性降低，突变蛋白正常或部分减少，三维结构分析表明活性部位没有直接参与。我们假设这三个新的突变会影响Gal-A蛋白，导致构象功能障碍。当在1-脱氧半乳糖基米星(DGJ)伴侣的存在下测试COS-1细胞和患者淋巴细胞中过表达的突变蛋白时，p.G183A和p.Y216C系统显示出GAL-A酶活性和蛋白质稳定性增加，而p.C52Y没有反应。我们强调，遗传学、生化和功能研究有助于阐明在FD中检测到的错义遗传损害的后果。ERT是Fabry患者的选择性治疗方法，但并不总是可能在所有涉及的器官中释放这种酶的活性形式。我们的研究支持这样的假设，即一种活性部位特异性的化学伴侣，可以口服给药，可能在治疗Gal-A构象缺陷方面有效。
Fabry Disease (FD) is an X-linked multisystemic lysosomal disorder caused by mutations of α-galactosidase (GLA) gene. Only a few of the 450 genetic lesions identified so far have been characterised by in vitro expression studies. Thus the significance of newly identified GLA nucleotide variants sin FD patients which lead to α-galactosidase (GAL-A) amino acid substitutions or intronic changes can be uncertain. We identified three GLA mutations: c155G>A (p.C52Y), c548G>C (p.G183A), c647A>G (p.Y216C) in as many individuals (two male; one female), and performed in vitro expression studies and Western Blot analysis, in order to clarify their functional effects. Reduced GAL-A activity and normal or partially reduced mutant proteins were present in all overexpressed mutant systems in which, three-dimensional structural analysis showed that the active site was not directly involved. We hypothesize that the three new mutations affect the GAL-A protein, leading to conformational FD. When, mutant proteins overexpressed in COS-1 cells and in patients’ lymphocytes were tested in the presence of the 1-deoxygalactonojirimicin (DGJ) chaperone, the p.G183A and p.Y216C systems showed increased GAL-A enzyme activities and protein stabilisation, while p.C52Y was not responsive. We underline that genetic, biochemical and functional studies are helpful in clarifying the consequences of the missense genetic lesions detected in FD. ERT is the elective therapy for Fabry patients but it is not always possible to issue the enzyme’s active form in all involved organs. Our study endorses the hypothesis that an active-site-specific chemical chaperone, which could be administered orally, might be effective in treating GAL-A conformational defects.
登录
查看更多内容
影响因子：
4
作者：
Morrone, A;Cavicchi, C;Zammarchi, E
通讯作者：
Zammarchi, E
影响因子：
19.6
作者：
Nakao, S;Kodama, C;Desnick, RJ
通讯作者：
Desnick, RJ
影响因子：
4.2
作者：
Benjamin, E. R.;Flanagan, J. J.;Valenzano, K. J.
通讯作者：
Valenzano, K. J.
DOI：
10.1006/bbrc.1995.2416
发表时间：
1995-09-25
影响因子：
3.1
作者：
OKUMIYA, T;ISHII, S;SUZUKI, Y
通讯作者：
SUZUKI, Y
影响因子：
3.9
作者：
Shimotori, Masaaki;Maruyama, Hiroki;Gejyo, Fumitake
通讯作者：
Gejyo, Fumitake