Complex Assembly Behavior During the Encapsulation of Green Fluorescent Protein Analogs in Virus Derived Protein Capsules
Complex Assembly Behavior During the Encapsulation of Green Fluorescent Protein Analogs in Virus Derived Protein Capsules
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DOI：
10.1002/mabi.201000030
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发表时间：
2010-05-14
影响因子：
4.6
通讯作者：
Cornelissen, Jeroen J. L. M.
中科院分区：
文献类型：
--
作者：
Minten, Inge J.;Nolte, Roeland J. M.;Cornelissen, Jeroen J. L. M.
与溶液中的游离酶相比，封装在纳米容器中的酶是活细胞内条件的更好模型。将多种酶封装在豇豆褪绿斑驳病毒 (CCMV) 衣壳内的第一步是将增强型绿色荧光蛋白 (EGFP) 附着到 CCMV 衣壳蛋白上。然后将衣壳蛋白-EGFP 复合物与野生型衣壳蛋白 (wt CP) 以不同的比例共组装。在复合物与 wt CP 比率较高时，每个衣壳的 EGFP 数量减少而不是趋于平稳。我们认为这种意想不到的行为是由 pH 诱导的衣壳蛋白-EGFP 复合物分解以及复合物的浓度和比例依赖性二聚化引起的，使其部分无法掺入衣壳中。
Enzymes encapsulated in nanocontainers are a better model of the conditions inside a living cell than free enzymes in solution. In a first step toward the encapsulation of multiple enzymes inside the cowpea chlorotic mottle virus (CCMV) capsid, enhanced green fluorescent protein (EGFP) was attached to CCMV capsid proteins. The capsid protein-EGFP complex was then co-assembled with wild-type capsid protein (wt CP) in various ratios. At higher complex to wt CP ratios, the number of EGFP per capsid decreased instead of leveling off. We propose that this unexpected behavior is caused by pH-induced disassembly of the capsid protein-EGFP complex as well as by concentration and ratio dependent dimerization of the complex, making it partially unavailable for incorporation into the capsid.