Conformational changes in bacteriophage phi 29 connector prevents DNA-binding activity.
Conformational changes in bacteriophage phi 29 connector prevents DNA-binding activity.
复制标题
噬菌体 phi 29 连接器的构象变化会阻止 DNA 结合活性。
DOI：
10.1016/s0022-2836(05)80189-5
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发表时间：
1990
影响因子：
5.6
通讯作者：
Carrascosa,JL
中科院分区：
文献类型：
--
作者：
Herranz,L;Bordas,J;Towns-Andrews,E;Mendez,E;Usobiaga,P;Carrascosa,JL
在衍生自噬菌体29的限定系统中的体外DNA包装活性取决于连接蛋白p10的化学完整性。p10的蛋白水解切割使proheads对DNA包装无活性。一个类似的处理孤立的连接器废除了原生p10的DNA结合活性，但一般的形状和大小的连接器没有改变所揭示的电子显微镜。分析超离心表明，蛋白水解的连接器有一个较小的沉降系数，而透析后的蛋白水解的p10氨基酸分析证实，16和19个氨基酸的氨基和羧基末端的损失，分别。低角X-射线散射显示，蛋白水解后，由一个小的回转半径的减少和重组的连接器的圆柱形内部的远端域。一级序列中的切割位点的表征使我们能够提出DNA结合结构域在连接器模型中的位置。
In vitroDNA packaging activity in a defined system derived from bacteriophageØ29 depends upon the chemical integrity of the connector protein p10. Proteolytic cleavage of p10 rendered the proheads inactive for DNA packaging. A similar treatment on isolated connectors abolished the DNA-binding activity of the native p10, but the general shape and size of the connector was not changed as revealed by electron microscopy. Analytical ultracentrifugation showed that the proteolyzed connectors had a smaller sedimentation coefficient, while amino acid analysis after dialysis of the proteolyzed p10 confirmed the loss of 16 and 19 amino acids from the amino and carboxy termini, respectively. Low angle X-ray scattering revealed that proteolysis was followed by a small decrease in the radius of gyration and a reorganization of the distal domain of the cylindrical inner part of the connector. Characterization of the cleavage sites in the primary sequence allowed us to propose the location of the DNA-binding domain in the connector model.