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编码. 中的一种高尔基体定位的 UDP-半乳糖转运蛋白。

encodes a Golgi-localized UDP-galactose transporter in .

机构信息

Departament of Immunology, Biomedical Sciences Institute, University of São Paulo, São Paulo, São Paulo 05508-900, SP, Brazil.

Institute Carlos Chagas, Fiocruz Paraná, Curitiba 81350-010, PR, Brasil.

出版信息

Parasitology. 2019 Sep;146(11):1379-1386. doi: 10.1017/S0031182019000738. Epub 2019 Jun 21.

Abstract

Survival and infectivity of trypanosomatids rely on cell-surface and secreted glycoconjugates, many of which contain a variable number of galactose residues. Incorporation of galactose to proteins and lipids occurs along the secretory pathway from UDP-galactose (UDP-Gal). Before being used in glycosylation reactions, however, this activated sugar donor must first be transported across the endoplasmic reticulum and Golgi membranes by a specific nucleotide sugar transporter (NST). In this study, we identified an UDP-Gal transporter (named TcNST2 and encoded by the TcCLB.504085.60 gene) from Trypanosoma cruzi, the etiological agent of Chagas disease. TcNST2 was identified by heterologous expression of selected putative nucleotide sugar transporters in a mutant Chinese Hamster Ovary cell line. TcNST2 mRNA levels were detected in all T. cruzi life-cycle forms, with an increase in expression in axenic amastigotes. Confocal microscope analysis indicated that the transporter is specifically localized to the Golgi apparatus. A three-dimensional model of TcNST2 suggested an overall structural conservation as compared with members of the metabolite transporter superfamily and also suggested specific features that could be related to its activity. The identification of this transporter is an important step toward a better understanding of glycoconjugate biosynthesis and the role NSTs play in this process in trypanosomatids.

摘要

生存和感染力依赖于表面和分泌糖缀合物的锥体虫,其中许多包含数量不定的半乳糖残基。半乳糖与蛋白质和脂质的结合发生在从 UDP-半乳糖(UDP-Gal)沿分泌途径。然而,在用于糖基化反应之前,这种激活的糖供体必须首先由特定的核苷酸糖转运蛋白(NST)跨内质网和高尔基体膜运输。在这项研究中,我们从恰加斯病的病原体克氏锥虫中鉴定了一种 UDP-Gal 转运蛋白(命名为 TcNST2,由 TcCLB.504085.60 基因编码)。通过在突变中国仓鼠卵巢细胞系中异源表达选定的假定核苷酸糖转运蛋白来鉴定 TcNST2。在所有 T. cruzi 生命周期形式中都检测到 TcNST2 mRNA 水平,在无细胞阿米巴滋养体中表达增加。共焦显微镜分析表明该转运蛋白特异性定位于高尔基体。与代谢物转运蛋白超家族成员的三维模型相比,TcNST2 的结构总体保守,并且还提示了与活性相关的特定特征。鉴定这种转运蛋白是更好地理解糖缀合物生物合成以及 NST 在锥虫中该过程中作用的重要步骤。

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