New addresses on an addressable virus nanoblock: Uniquely reactive lys residues on cowpea mosaic virus
New addresses on an addressable virus nanoblock: Uniquely reactive lys residues on cowpea mosaic virus
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DOI：
10.1016/j.chembiol.2004.04.011
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发表时间：
2004-06-01
影响因子：
--
通讯作者：
Lin, TW
中科院分区：
文献类型：
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作者：
Chatterji, A;Ochoa, WF;Lin, TW
豇豆花叶病毒 (CPMV) 是一种强大的二十面体对称平台，成功用于附着各种分子底物，包括蛋白质、荧光标记和金属。附着分子的对称分布和高局部浓度为 30 nm 颗粒带来了新颖的特性。我们报告了通过用精氨酸残基系统替换表面赖氨酸而产生的新 CPMV 试剂颗粒。确定天然颗粒上每个赖氨酸的相对反应性，然后通过用精氨酸残基替换所有其他赖氨酸，将两个最具反应性的赖氨酸残基创建为单个附着位点。金衍生化的结构分析不仅证实了这些独特的赖氨酸残基的特异性反应性，而且证明了它们显着不同的呈现环境。与定点胱氨酸突变相结合，现在可以对 CPMV 进行独特的双标记，从而扩大其作为可寻址纳米块的用途。
Cowpea mosaic virus (CPMV) is a robust, icosahedrally symmetric platform successfully used for attaching a variety of molecular substrates including proteins, fluorescent labels, and metals. The symmetric distribution and high local concentration of the attached molecules generates novel properties for the 30 nm particles. We report new CPMV reagent particles generated by systematic replacement of surface lysines with arginine residues. The relative reactivity of each lysine on the native particle was determined, and the two most reactive lysine residues were then created as single attachment sites by replacing all other lysines with arginine residues. Structural analysis of gold derivatization not only corroborated the specific reactivity of these unique lysine residues but also demonstrated their dramatically different presentation environment. Combined with site-directed cystine mutations, it is now possible to uniquely double label CPMV, expanding its use as an addressable nanoblock.