Power to the protein: enhancing and combining activities using the Spy toolbox.
Power to the protein: enhancing and combining activities using the Spy toolbox.
复制标题
DOI：
10.1039/d0sc01878c
复制
发表时间：
2020-07-28
期刊：
影响因子：
8.4
通讯作者：
Howarth M
中科院分区：
文献类型：
--
作者：
Keeble AH;Howarth M
具有简单和选择性反应性的肽扩展了蛋白质的功能，从单分子分析到潜在的临床应用。蛋白质的形状、大小和功能范围非常广泛。因此，需要通用的方法来克服这种多样性和流线蛋白质分析或应用。在这里，我们回顾SpyTag技术，现在在数百个出版物或专利中使用，以及其检测和控制蛋白质行为的潜力。SpyTag在结合其蛋白质伴侣SpyCatcher时形成自发和不可逆的异肽键，其中两个部分都是遗传编码的。这一对的新变体允许以接近扩散极限的速率进行反应，而可逆版本允许纯化SpyTagged蛋白或细胞内的调谐动态相互作用。SpyTag连接的蛋白质已经被建立到不同的纳米颗粒或表面，包括金，石墨烯和空气/水界面。SpyTag/SpyCatcher具有机械稳定性，因此被广泛用于研究蛋白质折叠和力敏感性。支架工具箱允许SpyTag融合物组装成定义的多聚体，从二聚体到180聚体，或无限的1D，2D或3D网络。目前正在评估二十面体多聚体用于疟疾、艾滋病毒和癌症的疫苗接种。对于酶，Spy技术增加了弹性，促进了底物通道，并组装了用于连续流动生物催化的水凝胶。通过抗体、发光二极管或病毒载体的模块化衍生，已经实现了功能的组合增加。在活细胞中，SpyTag允许对蛋白质运输进行成像，重新靶向CAR-T细胞杀伤，研究心脏收缩和控制核小体位置。简单的遗传编码和快速的不可逆反应为增强蛋白质功能提供了多种机会。我们描述了局限性以及未来的方向。
A peptide with simple and selective reactivity expands the function of proteins, from single molecule analysis to potential clinical application. Proteins span an extraordinary range of shapes, sizes and functionalities. Therefore generic approaches are needed to overcome this diversity and stream-line protein analysis or application. Here we review SpyTag technology, now used in hundreds of publications or patents, and its potential for detecting and controlling protein behaviour. SpyTag forms a spontaneous and irreversible isopeptide bond upon binding its protein partner SpyCatcher, where both parts are genetically-encoded. New variants of this pair allow reaction at a rate approaching the diffusion limit, while reversible versions allow purification of SpyTagged proteins or tuned dynamic interaction inside cells. Anchoring of SpyTag-linked proteins has been established to diverse nanoparticles or surfaces, including gold, graphene and the air/water interface. SpyTag/SpyCatcher is mechanically stable, so is widely used for investigating protein folding and force sensitivity. A toolbox of scaffolds allows SpyTag-fusions to be assembled into defined multimers, from dimers to 180-mers, or unlimited 1D, 2D or 3D networks. Icosahedral multimers are being evaluated for vaccination against malaria, HIV and cancer. For enzymes, Spy technology has increased resilience, promoted substrate channelling, and assembled hydrogels for continuous flow biocatalysis. Combinatorial increase in functionality has been achieved through modular derivatisation of antibodies, light-emitting diodes or viral vectors. In living cells, SpyTag allowed imaging of protein trafficking, retargeting of CAR-T cell killing, investigation of heart contraction, and control of nucleosome position. The simple genetic encoding and rapid irreversible reaction provide diverse opportunities to enhance protein function. We describe limitations as well as future directions.
登录
查看更多内容
影响因子：
7.3
作者：
Brune KD;Howarth M
通讯作者：
Howarth M
影响因子：
3.8
作者：
Botyanszki, Zsofia;Tay, Pei Kun R.;Joshi, Neel S.
通讯作者：
Joshi, Neel S.
DOI：
10.1073/pnas.1902163116
发表时间：
2019-05-28
影响因子：
11.1
作者：
Bartsch, Tobias F.;Hengel, Felicitas E.;Hudspeth, A. J.
通讯作者：
Hudspeth, A. J.
影响因子：
17.1
作者：
Bruun TUJ;Andersson AC;Draper SJ;Howarth M
通讯作者：
Howarth M
影响因子：
7.4
作者：
Anderson, George P.;Liu, Jinny L.;Goldman, Ellen R.
通讯作者：
Goldman, Ellen R.