Approaches to Identify and Characterise MYO6-Cargo Interactions.
Approaches to Identify and Characterise MYO6-Cargo Interactions.
复制标题
识别和表征 MYO6-货物相互作用的方法。
DOI：
10.1007/978-3-030-38062-5_15
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发表时间：
2020
影响因子：
--
通讯作者：
O'Loughlin T
中科院分区：
文献类型：
--
作者：
O'Loughlin T
考虑到肌动蛋白细胞骨架和相关肌球蛋白马达宿主的普遍性和重要性，发现它们与过多的细胞功能和病理学有关并不奇怪。虽然我们对肌球蛋白马达的生物物理性质的理解得到了其马达结构域的高度保守性和对肌球蛋白在骨骼肌收缩中的广泛研究的帮助，但我们对非肌肉肌球蛋白如何参与如此广泛的细胞过程的理解还不太清楚。现在已经确定，高度可变的肌球蛋白尾部负责将这些肌球蛋白靶向不同的细胞位点以实现特定功能，尽管已经鉴定了许多衔接蛋白，但我们目前对所涉及的细胞过程的理解相当有限。此外，随着越来越多的衔接蛋白，货物和复合物被确定，阐明所涉及的调控机制的重要性是必不可少的。Ca 2+，以及现在的磷酸化和泛素化，正在成为货物结合的重要调节因子，并且可能还涉及其他翻译后修饰。在肌球蛋白VI（MYO 6）的情况下，已经使用传统方法如酵母双杂交筛选和基于亲和力的下拉（pull-down）鉴定了许多直接结合配偶体。然而，这些方法仅成功鉴定了货物衔接子，而不是货物本身，其通常可能包含多蛋白复合物。此外，马达-适配器-货物相互作用本质上是动态的，并且通常是弱的、瞬时的和高度调节的，因此难以使用传统的基于亲和力的方法来捕获。在本章中，我们将讨论各种方法，包括功能蛋白质组学，已被用来发现和发现新的MYO 6相关蛋白和复合物，以及这项工作如何有助于更全面地了解这种独特的肌球蛋白马达的靶向和功能。
Given the prevalence and importance of the actin cytoskeleton and the host of associated myosin motors, it comes as no surprise to find that they are linked to a plethora of cellular functions and pathologies. Although our understanding of the biophysical properties of myosin motors has been aided by the high levels of conservation in their motor domains and the extensive work on myosin in skeletal muscle contraction, our understanding of how the nonmuscle myosins participate in such a wide variety of cellular processes is less clear. It is now well established that the highly variable myosin tails are responsible for targeting these myosins to distinct cellular sites for specific functions, and although a number of adaptor proteins have been identified, our current understanding of the cellular processes involved is rather limited. Furthermore, as more adaptor proteins, cargoes and complexes are identified, the importance of elucidating the regulatory mechanisms involved is essential. Ca2+, and now phosphorylation and ubiquitination, are emerging as important regulators of cargo binding, and it is likely that other post-translational modifications are also involved. In the case of myosin VI (MYO6), a number of immediate binding partners have been identified using traditional approaches such as yeast two-hybrid screens and affinity-based pull-downs. However, these methods have only been successful in identifying the cargo adaptors, but not the cargoes themselves, which may often comprise multi-protein complexes. Furthermore, motor-adaptor-cargo interactions are dynamic by nature and often weak, transient and highly regulated and therefore difficult to capture using traditional affinity-based methods. In this chapter we will discuss the various approaches including functional proteomics that have been used to uncover and characterise novel MYO6-associated proteins and complexes and how this work contributes to a fuller understanding of the targeting and function(s) of this unique myosin motor.
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DOI：
10.1146/annurev-cellbio-100809-151502
发表时间：
2011
影响因子：
11.3
作者：
Hartman MA;Finan D;Sivaramakrishnan S;Spudich JA
通讯作者：
Spudich JA
ErbB2直接激活交换因子Dock7促进雪旺细胞迁移。
DOI：
10.1083/jcb.200709033
发表时间：
2008-04-21
期刊：
The Journal of cell biology
影响因子：
--
作者：
Yamauchi J;Miyamoto Y;Chan JR;Tanoue A
通讯作者：
Tanoue A
DOI：
10.1073/pnas.91.14.6549
发表时间：
1994-07-05
影响因子：
11.1
作者：
BEMENT, WM;HASSON, T;MOOSEKER, MS
通讯作者：
MOOSEKER, MS
影响因子：
4.8
作者：
Yildiz, A;Park, H;Sweeney, HL
通讯作者：
Sweeney, HL
影响因子：
4.8
作者：
Shiba, Y;Katoh, Y;Nakayama, K
通讯作者：
Nakayama, K