Immobilization of diverse foreign proteins in viral polyhedra and potential application for protein microarrays
Immobilization of diverse foreign proteins in viral polyhedra and potential application for protein microarrays
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DOI：
10.1002/pmic.200500022
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发表时间：
2006-01-01
期刊：
影响因子：
3.4
通讯作者：
Mori, H
中科院分区：
文献类型：
--
作者：
Ikeda, K;Nakazawa, H;Mori, H
Cypoviruses是一种昆虫病毒，产生一种称为多面体的细胞质晶体颗粒，其中子代病毒粒子被封闭。病毒粒子结构蛋白VP 3与病毒粒子包埋成多面体有关。在这项研究中，我们确定了VP 3的氨基酸序列所需的病毒颗粒进入多面体的闭塞，并提出，这个序列可以作为一个固定化信号，以指导稳定掺入到多面体的外源蛋白。一项大规模的调查显示，固定信号实际上可以指导各种人类蛋白质并入多面体。免疫反应性和蛋白质-蛋白质相互作用的检测上含有固定化的外源蛋白质的多面体的表面上，这些颗粒被证明是高度稳定的抗脱水。我们表明，这些颗粒可以通过标准的点样和激光操作方法排列在载玻片上。因此，这种方法非常适合于蛋白质表达、纯化和蛋白质微阵列的开发。
Cypoviruses are insect viruses that produce a cytoplasmic crystalline particle called the polyhedron in which progeny virions are occluded. The virion structural protein, VP3, is implicated in the occlusion of viral particles into polyhedra. In this study, we determined the amino acid sequence of VP3 required for occlusion of viral particles into polyhedra and proposed that this sequence could be used as an immobilization signal to direct the stable incorporation of foreign proteins into polyhedra. A large-scale survey revealed that the immobilization signal could, in fact, direct the incorporation of a variety of human proteins into polyhedra. Immune reactivity and protein-protein interactions were detected on the surface of polyhedra containing immobilized foreign proteins, and these particles were shown to be highly stabilized against dehydration. We showed that these particles could be arrayed onto a glass slide by standard spotting and laser manipulation methods. Thus, this approach is well suited for protein expression, purification, and the development of protein microarrays.