Driedonks R A, Krijgsman P C, Mellema J E
Eur J Biochem. 1978 Jan 16;82(2):405-17. doi: 10.1111/j.1432-1033.1978.tb12035.x.
The polymerization of alfalfa mosaic virus (AMV) protein in the presence of homologous nucleic acids and a number of other natural and synthetic nucleic acids was studied. The conditions for optimal assembly were found to be pH 6.0 and low ionic strength (I = 0.1 M) at room temperature, irrespective of the type of nucleic acid. The resulting nucleoprotein particles exhibited the same structural characteristics as the virus. This information emerged from optical diffraction and computer filtering of electron micrographs from the reconstituted particles. Irrespective of the type of nucleic acid present the polymerization of the protein resulting in a nucleoprotein particle is a cooperative process. Evidence for this was obtained by nitrocellulose filter binding assay, sodium dodecylsulphate/polyacrylamide gel electrophoresis, sedimentation velocity and electron microscopy of the reaction mixtures. The rates and efficiencies of reconstitution were of the same order of magnitude for a number of ribonucleic acids. Sedimentation data derived from AMV protein and AMV RNA mixtures suggested the existence of a specific nucleation product in the first stage of assembly. The results are discussed in terms of a tentative model of the assembly, in which at least two different steps (nucleation and elongation) can be distinguished, each characterized by an association constant.
研究了苜蓿花叶病毒（AMV）蛋白在同源核酸以及多种其他天然和合成核酸存在下的聚合作用。发现最佳组装条件为室温下pH 6.0和低离子强度（I = 0.1 M），与核酸类型无关。所得核蛋白颗粒呈现出与病毒相同的结构特征。该信息来自对重组颗粒电子显微照片的光学衍射和计算机滤波。无论存在何种类型的核酸，导致核蛋白颗粒形成的蛋白质聚合都是一个协同过程。通过硝酸纤维素滤膜结合测定、十二烷基硫酸钠/聚丙烯酰胺凝胶电泳、沉降速度以及反应混合物的电子显微镜观察获得了相关证据。对于多种核糖核酸，重组的速率和效率处于同一数量级。源自AMV蛋白和AMV RNA混合物的沉降数据表明在组装的第一阶段存在特定的成核产物。根据一个初步的组装模型对结果进行了讨论，在该模型中至少可以区分两个不同的步骤（成核和延伸），每个步骤都有一个缔合常数作为特征。
