Elucidating the role of C-terminal post-translational modifications using protein semisynthesis strategies: α-synuclein phosphorylation at tyrosine 125.
Elucidating the role of C-terminal post-translational modifications using protein semisynthesis strategies: α-synuclein phosphorylation at tyrosine 125.
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DOI：
10.1021/ja210866j
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发表时间：
2012-03-21
影响因子：
15
通讯作者：
Lashuel, Hilal A.
中科院分区：
文献类型：
--
作者：
Hejjaoui, Mirva;Butterfield, Sara;Fauvet, Bruno;Vercruysse, Filip;Cui, Jia;Dikiy, Igor;Prudent, Michel;Olschewski, Diana;Zhang, Yan;Eliezer, David;Lashuel, Hilal A.
尽管越来越多的证据支持不同的翻译后修饰（PTM）在调节α-突触核蛋白（α-syn）聚集和毒性中的作用，但对每种修饰的功能后果以及这些修饰是否相互调节知之甚少。这种知识的缺乏主要是由于目前缺乏在α-syn中特定位点引入PTM的工具和方法。更具体地说，介导α-syn的C-末端酪氨酸残基的选择性和有效磷酸化的激酶仍有待鉴定。与磷酸丝氨酸和磷酸苏氨酸残基不同，在某些情况下可以通过丝氨酸/苏氨酸→谷氨酸或天冬氨酸取代来模拟，没有天然氨基酸可以模拟磷酸酪氨酸。为了解决这些挑战，我们开发了一种通用且有效的半合成策略，该策略能够位点特异性地引入单个或多个PTM，并以毫克量制备均匀的C-末端修饰形式的α-syn。这些进展使我们能够首次研究Y125的选择性磷酸化对α-syn的结构、聚集、膜结合和亚细胞定位的影响。用于位点特异性引入单个或PTM的半合成方法的发展代表了确定这种修饰在健康和疾病中的α-syn结构、聚集和功能中的作用的重要进展。
Despite increasing evidence that supports the role of different post-translational modifications (PTMs) in modulating α-synuclein (α-syn) aggregation and toxicity, relatively little is known about the functional consequences of each modification and whether or not these modifications are regulated by each other. This lack of knowledge arises primarily from the current lack of tools and methodologies for the site-specific introduction of PTMs in α-syn. More specifically, the kinases that mediate selective and efficient phosphorylation of C-terminal tyrosine residues of α-syn remain to be identified. Unlike phospho-serine and phospho-threonine residues, which in some cases can be mimicked by serine/threonine → glutamate or aspartate substitutions, there are no natural amino acids that can mimic phosphor-tyrosine. To address these challenges, we developed a general and efficient semisynthetic strategy that enables the site-specific introduction of single or multiple PTMs and the preparation of homogeneously C-terminal modified forms of α-syn in milligram quantities. These advances have allowed us to investigate, for the first time, the effects of selective phosphorylation at Y125 on the structure, aggregation, membrane binding and subcellular localization of α-syn. The development of semisynthetic methods for the site-specific introduction of single or PTMs represents an important advance toward determining the roles of such modifications in α-syn structure, aggregation and functions in heath and disease.
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影响因子：
4.1
作者：
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通讯作者：
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DOI：
10.1073/pnas.0711053105
发表时间：
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