Hyperstable De Novo Protein with a Dimeric Bisecting Topology.,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 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.)

Hyperstable De Novo Protein with a Dimeric Bisecting Topology.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-01-27 , DOI: 10.1021/acssynbio.9b00501
Naoya Kimura ₁ , Kenji Mochizuki _{2,

3} , Koji Umezawa _{4,

5} , Michael H Hecht ₆ , Ryoichi Arai _{1,

7,

8}

Affiliation

Recently, we designed and assembled protein nanobuilding blocks (PN-Blocks) from an intermolecularly folded dimeric de novo protein called WA20. Using this dimeric 4-helix bundle, we constructed a series of self-assembling supramolecular nanostructures including polyhedra and chain-type complexes. Here we describe the stabilization of WA20 by designing mutations that stabilize the helices and hydrophobic core. The redesigned proteins denature with substantially higher midpoints, with the most stable variant, called Super WA20 (SUWA), displaying an extremely high midpoint (Tm = 122 °C), much higher than the Tm of WA20 (75 °C). The crystal structure of SUWA reveals an intermolecularly folded dimer with bisecting U topology, similar to the parental WA20 structure, with two long α-helices of a protomer intertwined with the helices of another protomer. Molecular dynamics simulations demonstrate that the redesigned hydrophobic core in the center of SUWA significantly suppresses the deformation of helices observed in the same region of WA20, suggesting this is a critical factor stabilizing the SUWA structure. This hyperstable de novo protein is expected to be useful as nanoscale pillars of PN-Block components in new types of self-assembling nanoarchitectures.

中文翻译：

具有二聚对分拓扑的超稳定De Novo蛋白。

最近，我们从分子间折叠的二聚从头蛋白质WA20设计并组装了蛋白质纳米构件（PN-Blocks）。使用此二聚体4螺旋束，我们构建了一系列自组装的超分子纳米结构，包括多面体和链型复合物。在这里，我们通过设计使螺旋和疏水核稳定的突变来描述WA20的稳定。重新设计的蛋白质具有较高的中点变性，最稳定的变体称为Super WA20（SUWA），显示极高的中点（Tm = 122°C），远高于WA20的Tm（75°C）。SUWA的晶体结构显示了一个分子对折的二聚体，具有二等分的U形拓扑结构，类似于亲本WA20的结构，一个protomer的两个长α-螺旋与另一个protomer的螺旋缠绕在一起。分子动力学模拟表明，在SUWA中心重新设计的疏水核显着抑制了在WA20的同一区域中观察到的螺旋的变形，这表明这是稳定SUWA结构的关键因素。这种超稳定的从头蛋白有望用作新型自组装纳米结构中PN-Block组分的纳米级支柱。

更新日期：2020-01-27

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11