Rational Design of Phosphorylation-Responsive Coiled Coil-Peptide Assemblies.
Rational Design of Phosphorylation-Responsive Coiled Coil-Peptide Assemblies.
复制标题
DOI：
10.1021/acssynbio.3c00064
复制
发表时间：
2023-04-21
影响因子：
4.7
通讯作者：
Woolfson, Derek N.
中科院分区：
文献类型：
--
作者：
Thompson, Harry F.;Beesley, Joseph L.;Langlands, Hannah D.;Edgell, Caitlin L.;Savery, Nigel J.;Woolfson, Derek N.
关键词：
响应化学和物理线索而改变状态的从头肽和蛋白质将推进蛋白质设计和合成生物学。在这里，我们报告了两种设计的系统，它们分别在位点特异性磷酸化和去磷酸化时分解和重组。作为起点，我们使用超热稳定从头反平行和平行卷曲螺旋异四聚体（即 A2B2 系统）来提供下游应用的控制。通过添加蛋白激酶 A 磷酸化位点 R-R-X-S 来整合开关，并放置磷酸受体丝氨酸残基以最大限度地破坏卷曲螺旋界面。未磷酸化的肽按照设计组装，并在加热时可逆地展开。在组装状态中添加激酶可将其展开，半衰期≤5 分钟。磷酸化被 Lambda 蛋白磷酸酶逆转，导致四聚体重新组装。我们设想，新设计的卷绕线圈组件、开关及其机械模型将在合成生物学、生物材料和生物技术应用中有用。
De novo peptides and proteins that switch state in response to chemical and physical cues would advance protein design and synthetic biology. Here we report two designed systems that disassemble and reassemble upon site-specific phosphorylation and dephosphorylation, respectively. As starting points, we use hyperthermostable de novo antiparallel and parallel coiled-coil heterotetramers, i.e., A2B2 systems, to afford control in downstream applications. The switches are incorporated by adding protein kinase A phosphorylation sites, R-R-X-S, with the phosphoacceptor serine residues placed to maximize disruption of the coiled-coil interfaces. The unphosphorylated peptides assemble as designed and unfold reversibly when heated. Addition of kinase to the assembled states unfolds them with half-lives of ≤5 min. Phosphorylation is reversed by Lambda Protein Phosphatase resulting in tetramer reassembly. We envisage that the new de novo designed coiled-coil components, the switches, and a mechanistic model for them will be useful in synthetic biology, biomaterials, and biotechnology applications.
登录
查看更多内容
影响因子：
3.2
作者：
Gao, Feng;Thornley, Blair S.;Zondlo, Neal J.
通讯作者：
Zondlo, Neal J.
影响因子：
5.6
作者：
Grigoryan G;Degrado WF
通讯作者：
Degrado WF
影响因子：
15
作者：
Cristie-Dayid, Ajitha S.;Chen, Junjie;Marsh, E. Neil G.
通讯作者：
Marsh, E. Neil G.
影响因子：
3.2
作者：
Aupic, Jana;Lapenta, Fabio;Jerala, Roman
通讯作者：
Jerala, Roman
影响因子：
56.9
作者：
Chen, Zibo;Kibler, Ryan D.;Baker, David
通讯作者：
Baker, David