Akhter Tahmina, Zhao Li, Kohda Atsushi, Mio Kazuhiro, Kanamaru Shuji, Arisaka Fumio
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4359-B39 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Biochim Biophys Acta. 2007 Aug;1774(8):1036-43. doi: 10.1016/j.bbapap.2007.05.011. Epub 2007 May 29.
After packaging of DNA into the head of bacteriophage T4 is completed, a neck is formed at the portal vertex of the head to be ready for the tail attachment. The main components of the neck are gp13 and gp14 (gp: gene product), which consist of 309 and 256 amino acid residues, respectively. In order to elucidate the structure and subunit arrangement in the neck, overexpression systems of gene 13 and gene 14 were constructed and purified to homogeneity. Far-UV circular dichroism (CD) spectra of gp13 and gp14 indicated that gp13 is rich in alpha-helices whereas gp14 is rich in beta-sheets. Sedimentation velocity analysis of gp13 and gp14 revealed that both proteins are present as monomers in solution. The frictional ratios (f/f(0)) of the two proteins indicated that gp14 has a more elongated shape than gp13. Although isolated gp13 and gp14 do not interact with each other when mixed under physiological conditions, they form a hetero-oligomer complex with the stoichiometry of 10:5 after treatment with ammonium sulfate. Electron microscopy of this complex has shown that it forms a ring-like structure of 15 nm in diameter.
将DNA包装到噬菌体T4头部完成后,在头部的门户顶点处形成颈部,为尾部附着做好准备。颈部的主要成分是gp13和gp14(gp:基因产物),它们分别由309和256个氨基酸残基组成。为了阐明颈部的结构和亚基排列,构建了基因13和基因14的过表达系统,并纯化至均一。gp13和gp14的远紫外圆二色性(CD)光谱表明,gp13富含α-螺旋,而gp14富含β-折叠。gp13和gp14的沉降速度分析表明,这两种蛋白质在溶液中均以单体形式存在。两种蛋白质的摩擦系数(f/f(0))表明,gp14的形状比gp13更细长。尽管在生理条件下混合时,分离的gp13和gp14不会相互作用,但用硫酸铵处理后,它们会形成化学计量比为10:5的异源寡聚体复合物。该复合物的电子显微镜观察表明,它形成了直径为15 nm的环状结构。