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Chemical reactivity of brome mosaic virus capsid protein

机译：溴化花叶病毒衣壳蛋白的化学反应活性

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Viral particles are biological machines that have evolved to package, protect, and deliver the viral genome into the host via regulated conformational changes of virions. We have developed a procedure to modify lysine residues with S-methylthioacetimidate across the pH range from 5.5 to 8.5. Lysine residues that are not completely modified are involved in tertiary or quaternary structural interactions, and their extent of modification can be quantified as a function of pH. This procedure was applied to the pH-dependent structural transitions of brome mosaic virus (BMV). As the reaction pH increases from 5.5 to 8.5, the average number of modified lysine residues in the BMV capsid protein increases from 6 to 12, correlating well with the known pH-dependent swelling behavior of BMV virions. The extent of reaction of each of the capsid protein's lysine residues has been quantified at eight pH values using coupled liquid chromatography-tandem mass spectrometry. Each lysine can be assigned to one of three structural classes identified by inspection of the BMV virion crystal structure. Several lysine residues display reactivity that indicates their involvement in dynamic interactions that are not obvious in the crystal structure. The influence of several capsid protein mutants on the pH-dependent structural transition of BMV has also been investigated. Mutant H75Q exhibits an altered swelling transition accompanying solution pH increases. The H75Q capsids show increased reactivity at lysine residues 64 and 130, residues distal from the dimer interface occupied by H75, across the entire pH range.
机译：病毒颗粒是通过调节病毒体的构象变化，将病毒基因组包装，保护并传递到宿主中的生物机器。我们已经开发出一种程序，可以在5.5至8.5的pH范围内用S-甲基硫代乙酰基亚氨酸修饰赖氨酸残基。未完全修饰的赖氨酸残基参与三级或四级结构相互作用，其修饰程度可根据pH进行量化。该程序应用于溴代花叶病毒（BMV）的pH依赖性结构转变。随着反应pH从5.5增加到8.5，BMV衣壳蛋白中修饰的赖氨酸残基的平均数从6增加到12，与BMV病毒体的已知pH依赖性溶胀行为良好相关。每个衣壳蛋白赖氨酸残基的反应程度已使用液相色谱-串联质谱在8个pH值进行了定量。可以将每个赖氨酸分配给通过检查BMV病毒体晶体结构确定的三个结构类别之一。几个赖氨酸残基显示出反应性，表明它们参与了晶体结构中不明显的动态相互作用。还研究了几种衣壳蛋白突变体对BMV pH依赖性结构转变的影响。突变体H75Q随溶液pH值的增加而呈现出变化的溶胀转变。在整个pH范围内，H75Q衣壳在赖氨酸残基64和130（H75占据的二聚体界面远端的残基）处显示出更高的反应性。

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