Quantitative characterization of all single amino acid variants of a viral capsid-based drug delivery vehicle.
Quantitative characterization of all single amino acid variants of a viral capsid-based drug delivery vehicle.
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DOI：
10.1038/s41467-018-03783-y
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发表时间：
2018-04-11
影响因子：
16.6
通讯作者：
Tullman-Ercek D
中科院分区：
文献类型：
--
作者：
Hartman EC;Jakobson CM;Favor AH;Lobba MJ;Álvarez-Benedicto E;Francis MB;Tullman-Ercek D
自组装蛋白质对生物系统和工业技术至关重要，但预测突变如何影响自组装仍然是一个重大挑战。在这里，我们报告了一种技术，称为SyMAPS（系统突变和组装粒子选择），可用于表征自组装病毒结构蛋白的所有单个氨基酸变体的组装能力。对MS2噬菌体外壳蛋白的SyMAPS研究揭示了一个高分辨率的适应性景观，挑战了蛋白质工程的一些传统假设。另一轮选择鉴定了先前未知的变体（CP[T71H]），其在中性pH下稳定，但对酸性条件的耐受性低于野生型外壳蛋白。由这种变体形成的衣壳可能更易于在晚期内体或早期溶酶体中解体，这是一种有利于递送应用的特征。除了提供病毒样颗粒的突变蓝图，SyMAPS可以很容易地应用于其他自组装蛋白质。自组装蛋白在工业技术中有重要的应用，但很难预测突变如何影响组装。在这里，作者介绍了SyMAPS，耦合全面的密码子诱变与高通量测序，并将其应用于噬菌体衣壳蛋白。
Self-assembling proteins are critical to biological systems and industrial technologies, but predicting how mutations affect self-assembly remains a significant challenge. Here, we report a technique, termed SyMAPS (Systematic Mutation and Assembled Particle Selection), that can be used to characterize the assembly competency of all single amino acid variants of a self-assembling viral structural protein. SyMAPS studies on the MS2 bacteriophage coat protein revealed a high-resolution fitness landscape that challenges some conventional assumptions of protein engineering. An additional round of selection identified a previously unknown variant (CP[T71H]) that is stable at neutral pH but less tolerant to acidic conditions than the wild-type coat protein. The capsids formed by this variant could be more amenable to disassembly in late endosomes or early lysosomes—a feature that is advantageous for delivery applications. In addition to providing a mutability blueprint for virus-like particles, SyMAPS can be readily applied to other self-assembling proteins. Self-assembling proteins have important applications in industrial technologies, but it is difficult to predict how mutations affect assembly. Here the authors present SyMAPS, coupling comprehensive codon mutagenesis with high-throughput sequencing, and apply it to bacteriophage capsid protein.
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影响因子：
17.1
作者：
Glasgow, Jeff E.;Capehart, Stacy L.;Francis, Matthew B.;Tullman-Ercek, Danielle
通讯作者：
Tullman-Ercek, Danielle
影响因子：
17.1
作者：
Ashley CE;Carnes EC;Phillips GK;Durfee PN;Buley MD;Lino CA;Padilla DP;Phillips B;Carter MB;Willman CL;Brinker CJ;Caldeira Jdo C;Chackerian B;Wharton W;Peabody DS
通讯作者：
Peabody DS
影响因子：
32.4
作者：
Ferguson AL;Mann JK;Omarjee S;Ndung'u T;Walker BD;Chakraborty AK
通讯作者：
Chakraborty AK
影响因子：
4.9
作者：
Carrico, Zachary M.;Romanini, Dante W.;Francis, Matthew B.
通讯作者：
Francis, Matthew B.
影响因子：
10.7
作者：
Firnberg E;Labonte JW;Gray JJ;Ostermeier M
通讯作者：
Ostermeier M