The energetic contributions of scaffolding and coat proteins to the assembly of bacteriophage procapsids.
The energetic contributions of scaffolding and coat proteins to the assembly of bacteriophage procapsids.
复制标题
支架和外壳蛋白对噬菌体原衣壳组装的能量贡献。
DOI：
10.1016/j.virol.2012.03.017
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Teschke,CarolynM
中科院分区：
文献类型：
--
作者：
Zlotnick,Adam;Suhanovsky,MargaretM;Teschke,CarolynM
噬菌体P22原囊体的体外组装需要外壳蛋白和亚化学计量浓度的内部支架蛋白。如果没有支架蛋白，外壳蛋白会异常组装，但只有在较高浓度时才会异常组装。过多的支架蛋白导致部分原壳。通过将原衣壳视为一个可以结合并被支架蛋白稳定的晶格，我们剖析了原衣壳组装作为蛋白质浓度和支架/外壳蛋白比的函数。我们观察到：(i)与异常聚合物形成相比，原衣壳的衣壳结合更弱；（ii）支架蛋白对原衣壳的稳定性贡献虽小，但足以支持原衣壳的形成；（iii）有多种类型的支架蛋白结合位点。这种方法应该适用于其他异质病毒组装反应，并将促进我们操纵这种体外反应以组装探针和开发纳米颗粒的能力。
In vitro assembly of bacteriophage P22 procapsids requires coat protein and sub-stoichiometric concentrations of the internal scaffolding protein. If there is no scaffolding protein, coat protein assembles aberrantly, but only at higher concentrations. Too much scaffolding protein results in partial procapsids. By treating the procapsid as a lattice that can bind and be stabilized by scaffolding protein we dissect procapsid assembly as a function of protein concentration and scaffolding/coat protein ratio. We observe that (i) the coat–coat association is weaker for procapsids than for aberrant polymer formation, (ii) scaffolding protein makes a small but sufficient contribution to stability to favor the procapsid form, and (iii) there are multiple classes of scaffolding protein binding sites. This approach should be applicable to other heterogeneous virus assembly reactions and will facilitate our ability to manipulate such in vitro reactions to probe assembly, and for development of nanoparticles.