Modification of bacterial microcompartments with target biomolecules via post-translational SpyTagging.
Modification of bacterial microcompartments with target biomolecules via post-translational SpyTagging.
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细菌微室（BMCs）是细菌内部形成的蛋白质类细胞器结构，通常包裹酶和细胞过程，特别是允许有毒中间体免受一般细胞环境的影响。除了它们的生物学作用外，通过表面修饰，它们还可以作为潜在的药物载体和多价抗原展示支架。在这里，我们使用翻译后修饰方法，使用铜自由点击化学，将SpyTag附着在目标蛋白分子上，以附着在特定的SpyCatcher修饰的BMC壳蛋白上。我们证明翻译后的SpyTagged材料可以与SpyCatcher修饰的BMC发生反应，并在BMC表面显示其存在，从而使这些结构作为多价抗原展示支架用于疫苗开发的未来研究成为可能。这种翻译后的“点击”方法克服了对SpyTag进行遗传编码的必要性，避免了表达量的任何潜在降低，并扩大了SpyTag/SpyCatcher疫苗支架的范围，以形成肽表位疫苗和小分子递送剂。翻译后的策略为生产SpyTagged分子适用于间谍为基础的共轭描述。
Bacterial microcompartments (BMCs) are proteinaceous organelle-like structures formed within bacteria, often encapsulating enzymes and cellular processes, in particular, allowing toxic intermediates to be shielded from the general cellular environment. Outside of their biological role they are of interest, through surface modification, as potential drug carriers and polyvalent antigen display scaffolds. Here we use a post-translational modification approach, using copper free click chemistry, to attach a SpyTag to a target protein molecule for attachment to a specific SpyCatcher modified BMC shell protein. We demonstrate that a post-translationally SpyTagged material can react with a SpyCatcher modified BMC and show its presence on the surface of BMCs, enabling future investigation of these structures as polyvalent antigen display scaffolds for vaccine development. This post-translational ‘click’ methodology overcomes the necessity to genetically encode the SpyTag, avoids any potential reduction in expression yield and expands the scope of SpyTag/SpyCatcher vaccine scaffolds to form peptide epitope vaccines and small molecule delivery agents. A post-translational strategy for the production of SpyTagged molecules suitable for use in Spy based conjugations is described.
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影响因子：
3.7
作者：
Pryde DC;Jones LH;Gervais DP;Stead DR;Blakemore DC;Selby MD;Brown AD;Coe JW;Badland M;Beal DM;Glen R;Wharton Y;Miller GJ;White P;Zhang N;Benoit M;Robertson K;Merson JR;Davis HL;McCluskie MJ
通讯作者：
McCluskie MJ
影响因子：
3.1
作者：
Alam, Md. Kausar;El-Sayed, Ayman;Geyer, C. Ronald
通讯作者：
Geyer, C. Ronald
影响因子：
3.7
作者：
Beerli RR;Hell T;Merkel AS;Grawunder U
通讯作者：
Grawunder U
用于靶向酶定位的工程蛋白质纳米室。
影响因子：
3.7
作者：
Choudhary S;Quin MB;Sanders MA;Johnson ET;Schmidt-Dannert C
通讯作者：
Schmidt-Dannert C
DOI：
10.1073/pnas.1207516109
发表时间：
2012-09-11
影响因子：
11.1
作者：
Fan, Chenguang;Cheng, Shouqiang;Bobik, Thomas A.
通讯作者：
Bobik, Thomas A.