Design of functionalised circular tandem repeat proteins with longer repeat topologies and enhanced subunit contact surfaces.
Design of functionalised circular tandem repeat proteins with longer repeat topologies and enhanced subunit contact surfaces.
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具有较长重复拓扑结构和增强亚基接触表面的功能化环状串联重复蛋白的设计。
DOI：
10.1038/s42003-021-02766-y
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发表时间：
2021-10-29
影响因子：
5.9
通讯作者：
Stoddard BL
中科院分区：
文献类型：
--
作者：
Hallinan JP;Doyle LA;Shen BW;Gewe MM;Takushi B;Kennedy MA;Friend D;Roberts JM;Bradley P;Stoddard BL
环状串联重复蛋白（“cTRP”）是从头设计的蛋白质支架（在本研究和先前的研究中，基于反平行双螺旋束），其含有重复的蛋白质序列和结构基序并形成闭合的环状结构。它们可以显示出显著的稳定性和溶解性，宽范围的尺寸，并且可用作生物技术应用的蛋白质展示颗粒。然而，cTRP也表现出来自较小亚基的低效自组装。在这项研究中，我们描述了新一代的cTRP，具有更长的重复序列和增加的相互作用表面，这增强了两个显着不同大小的同源三聚体构建体的自组装。最后，我们用（1）肽结合SH 2结构域的六聚体阵列和（2）抗SARS CoV-2 VHH结构域的三聚体阵列证明了这些构建体的功能化。后者证明能够对病毒受体结合结构域具有亚纳摩尔结合亲和力和有效的病毒中和功能。Jazmine Hallinan等人报道了具有增强的自组装的新一代环状串联重复蛋白的开发。这些构建体与SARS CoV-2 VHH结构域的功能化导致对病毒受体结合结构域的亚纳摩尔结合亲和力。
Circular tandem repeat proteins (‘cTRPs’) are de novo designed protein scaffolds (in this and prior studies, based on antiparallel two-helix bundles) that contain repeated protein sequences and structural motifs and form closed circular structures. They can display significant stability and solubility, a wide range of sizes, and are useful as protein display particles for biotechnology applications. However, cTRPs also demonstrate inefficient self-assembly from smaller subunits. In this study, we describe a new generation of cTRPs, with longer repeats and increased interaction surfaces, which enhanced the self-assembly of two significantly different sizes of homotrimeric constructs. Finally, we demonstrated functionalization of these constructs with (1) a hexameric array of peptide-binding SH2 domains, and (2) a trimeric array of anti-SARS CoV-2 VHH domains. The latter proved capable of sub-nanomolar binding affinities towards the viral receptor binding domain and potent viral neutralization function. Jazmine Hallinan et al. report the development of a new generation of circular tandem repeat proteins with enhanced self-assembly. Functionalisation of these constructs with SARS CoV-2 VHH domains resulted in sub-nanomolar binding affinity to the viral receptor binding domain.
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影响因子：
4.7
作者：
Moore, Richard;Chandrahas, Anita;Bleris', Leonidas
通讯作者：
Bleris', Leonidas
影响因子：
6.7
作者：
Antanasijevic, Aleksandar;Ueda, George;Ward, Andrew B.
通讯作者：
Ward, Andrew B.
影响因子：
46.9
作者：
Castellana M;Wilson MZ;Xu Y;Joshi P;Cristea IM;Rabinowitz JD;Gitai Z;Wingreen NS
通讯作者：
Wingreen NS
影响因子：
21.8
作者：
Fallas JA;Ueda G;Sheffler W;Nguyen V;McNamara DE;Sankaran B;Pereira JH;Parmeggiani F;Brunette TJ;Cascio D;Yeates TR;Zwart P;Baker D
通讯作者：
Baker D
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH