当前位置：
X-MOL 学术
›
J. Am. Chem. Soc.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
An Atypical Mechanism of Split Intein Molecular Recognition and Folding
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2018-08-29 , DOI: 10.1021/jacs.8b07334 Adam J Stevens 1 , Giridhar Sekar 2 , Josef A Gramespacher 1 , David Cowburn 2 , Tom W Muir 1
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2018-08-29 , DOI: 10.1021/jacs.8b07334 Adam J Stevens 1 , Giridhar Sekar 2 , Josef A Gramespacher 1 , David Cowburn 2 , Tom W Muir 1
Affiliation
|
Split inteins associate to trigger protein splicing in trans, a post-translational modification in which protein sequences fused to the intein pair are ligated together in a traceless manner. Recently, a family of naturally split inteins has been identified that is split at a noncanonical location in the primary sequence. These atypically split inteins show considerable promise in protein engineering applications; however, the mechanism by which they associate is unclear and must be different from that of previously characterized canonically split inteins due to unique topological restrictions. Here, we use a consensus design strategy to generate an atypical split intein pair (Cat) that has greatly improved activity and is amenable to detailed biochemical and biophysical analysis. Guided by the solution structure of Cat, we show that the association of the fragments involves a disorder-to-order structural transition driven by hydrophobic interactions. This molecular recognition mechanism satisfies the topological constraints of the intein fold and, importantly, ensures that premature chemistry does not occur prior to fragment complementation. Our data lead a common blueprint for split intein complementation in which localized structural rearrangements are used to drive folding and regulate protein-splicing activity.
中文翻译：
分裂内含肽分子识别和折叠的非典型机制
分裂的内含肽与触发蛋白质反式剪接相关，这是一种翻译后修饰，其中融合到内含肽对的蛋白质序列以无痕方式连接在一起。最近，已鉴定出一个自然分裂的内含肽家族，其在一级序列中的非规范位置处分裂。这些非典型分裂内含子在蛋白质工程应用中显示出巨大的前景。然而，它们结合的机制尚不清楚，并且由于独特的拓扑限制，必定与之前描述的规范分裂内含肽的机制不同。在这里，我们使用共识设计策略来生成非典型分裂内含肽对 (Cat)，其活性大大提高，并且适合详细的生化和生物物理分析。在 Cat 溶液结构的指导下，我们表明片段的缔合涉及由疏水相互作用驱动的无序到有序的结构转变。这种分子识别机制满足内含肽折叠的拓扑限制，并且重要的是，确保在片段互补之前不会发生过早的化学反应。我们的数据引导了分裂内含肽互补的通用蓝图，其中局部结构重排用于驱动折叠和调节蛋白质剪接活性。
更新日期：2018-08-29
中文翻译：
分裂内含肽分子识别和折叠的非典型机制
分裂的内含肽与触发蛋白质反式剪接相关，这是一种翻译后修饰，其中融合到内含肽对的蛋白质序列以无痕方式连接在一起。最近，已鉴定出一个自然分裂的内含肽家族，其在一级序列中的非规范位置处分裂。这些非典型分裂内含子在蛋白质工程应用中显示出巨大的前景。然而，它们结合的机制尚不清楚，并且由于独特的拓扑限制，必定与之前描述的规范分裂内含肽的机制不同。在这里，我们使用共识设计策略来生成非典型分裂内含肽对 (Cat)，其活性大大提高，并且适合详细的生化和生物物理分析。在 Cat 溶液结构的指导下，我们表明片段的缔合涉及由疏水相互作用驱动的无序到有序的结构转变。这种分子识别机制满足内含肽折叠的拓扑限制，并且重要的是，确保在片段互补之前不会发生过早的化学反应。我们的数据引导了分裂内含肽互补的通用蓝图，其中局部结构重排用于驱动折叠和调节蛋白质剪接活性。
京公网安备 11010802027423号