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利用新型蛋白质生产方法挖掘微管蛋白的宝藏。

TAPping into the treasures of tubulin using novel protein production methods.

机构信息

Department of Cell Biology, Erasmus MC, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands.

Department of Cell Biology, Erasmus MC, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands

出版信息

Essays Biochem. 2018 Dec 7;62(6):781-792. doi: 10.1042/EBC20180033.

Abstract

Microtubules are cytoskeletal elements with important cellular functions, whose dynamic behaviour and properties are in part regulated by microtubule-associated proteins (MAPs). The building block of microtubules is tubulin, a heterodimer of α- and β-tubulin subunits. Longitudinal interactions between tubulin dimers facilitate a head-to-tail arrangement of dimers into protofilaments, while lateral interactions allow the formation of a hollow microtubule tube that mostly contains 13 protofilaments. Highly homologous α- and β-tubulin isotypes exist, which are encoded by multi-gene families. studies on microtubules and MAPs have largely relied on brain-derived tubulin preparations. However, these consist of an unknown mix of tubulin isotypes with undefined post-translational modifications. This has blocked studies on the functions of tubulin isotypes and the effects of tubulin mutations found in human neurological disorders. Fortunately, various methodologies to produce recombinant mammalian tubulins have become available in the last years, allowing researchers to overcome this barrier. In addition, affinity-based purification of tagged tubulins and identification of tubulin-associated proteins (TAPs) by mass spectrometry has revealed the 'tubulome' of mammalian cells. Future experiments with recombinant tubulins should allow a detailed description of how tubulin isotype influences basic microtubule behaviour, and how MAPs and TAPs impinge on tubulin isotypes and microtubule-based processes in different cell types.

摘要

微管是具有重要细胞功能的细胞骨架元件,其动态行为和特性部分受微管相关蛋白(MAPs)的调节。微管的结构单元是微管蛋白,它是α-和β-微管蛋白亚基的异二聚体。微管蛋白二聚体的纵向相互作用促进二聚体头尾排列成原纤维,而侧向相互作用允许形成一个含有 13 个原纤维的空心微管管。存在高度同源的α-和β-微管蛋白同工型,它们由多基因家族编码。对微管和 MAP 的研究在很大程度上依赖于脑源性微管蛋白制剂。然而,这些制剂由微管蛋白同工型的未知混合物组成,其翻译后修饰是未知的。这阻碍了对微管蛋白同工型功能的研究,以及对人类神经紊乱中发现的微管蛋白突变的研究。幸运的是,近年来已经出现了各种生产重组哺乳动物微管蛋白的方法,使研究人员能够克服这一障碍。此外,通过亲和纯化标记的微管蛋白和通过质谱鉴定微管相关蛋白(TAPs),揭示了哺乳动物细胞的“微管组”。未来使用重组微管蛋白的实验应该能够详细描述微管蛋白同工型如何影响基本的微管行为,以及 MAPs 和 TAPs 如何影响不同细胞类型中的微管蛋白同工型和基于微管的过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d78c/6281476/a35d20a5debb/ebc-62-ebc20180033-g1.jpg

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