Chorein, the protein responsible for chorea-acanthocytosis, interacts with β-adducin and β-actin
Chorein, the protein responsible for chorea-acanthocytosis, interacts with β-adducin and β-actin
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DOI：
10.1016/j.bbrc.2013.10.011
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发表时间：
2013-11-08
影响因子：
3.1
通讯作者：
Sano, Akira
中科院分区：
文献类型：
--
作者：
Shiokawa, Nari;Nakamura, Masayuki;Sano, Akira
舞蹈病-棘红细胞增多症（ChAc）是一种以纹状体神经变性和棘红细胞增多症为特征的常染色体隐性遗传性疾病，由液泡蛋白分选13同源物A（VPS 13 A）基因的功能缺失突变引起。VPS 13 A编码chorein，其在分子水平上的生理功能知之甚少。在这项研究中，我们表明，chorein与β-内收蛋白和β-肌动蛋白相互作用。我们首先使用蛋白质组学分析比较人红细胞膜中的蛋白质表达。β-内收蛋白亚型1和β-肌动蛋白的蛋白质水平显着降低，在红细胞膜从ChAc患者。随后的免疫共沉淀（co-IP）和反向co-IP测定使用从chorein过表达的人胚肾293（HEK 293）细胞的提取物，显示β-内收蛋白（亚型I和2）和β-肌动蛋白与chorein相互作用。使用chorein过表达的HEK 293细胞的免疫细胞化学分析证明chorein与β-内收蛋白和β-肌动蛋白共定位。此外，β-内收蛋白亚型1的免疫反应性在基因靶向ChAc模型小鼠的纹状体中显著降低。内收蛋白和肌动蛋白是细胞膜骨架蛋白，参与突触功能。β-内收蛋白的表达仅限于脑和造血组织，对应于ChAc的主要病理损害，从而暗示β-内收蛋白和β-肌动蛋白在ChAc发病机制中。(C)2013 Elsevier Inc. All rights reserved.
Chorea-acanthocytosis (ChAc) is an autosomal, recessive hereditary disease characterized by striatal neurodegeneration and acanthocytosis, and caused by loss of function mutations in the vacuolar protein sorting 13 homolog A (VPS13A) gene. VPS13A encodes chorein whose physiological function at the molecular level is poorly understood. In this study, we show that chorein interacts with beta-adducin and beta-actin. We first compare protein expression in human erythrocyte membranes using proteomic analysis. Protein levels of beta-adducin isoform 1 and beta-actin are markedly decreased in erythrocyte membranes from a ChAc patient. Subsequent co-immunoprecipitation (co-IP) and reverse co-IP assays using extracts from chorein-overexpressing human embryonic kidney 293 (HEK293) cells, shows that beta-adducin (isoforms I and 2) and beta-actin interact with chorein. Immunocytochemical analysis using chorein-overexpressing HEK293 cells demonstrates co-localization of chorein with beta-adducin and (beta-actin. In addition, immunoreactivity of beta-adducin isoform 1 is significantly decreased in the striatum of gene-targeted ChAc-model mice. Adducin and actin are membrane cytoskeletal proteins, involved in synaptic function. Expression of beta-adducin is restricted to the brain and hematopoietic tissues, corresponding to the main pathological lesions of ChAc, and thereby implicating beta-adducin and beta-actin in ChAc pathogenesis. (C) 2013 Elsevier Inc. All rights reserved.