Observation of the membrane binding activity and domain structure of gpV, which comprises the tail spike of bacteriophage P2.
Observation of the membrane binding activity and domain structure of gpV, which comprises the tail spike of bacteriophage P2.
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DOI：
10.1021/bi900928n
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发表时间：
2009-10
期刊：
影响因子：
2.9
通讯作者：
Y. Kageyama;M. Murayama;T. Onodera;S. Yamada;H. Fukada;M. Kudou;K. Tsumoto;Y. Toyama;Syunsaku Kado;K. Kubota;S. Takeda
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文献类型：
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作者：
Y. Kageyama;M. Murayama;T. Onodera;S. Yamada;H. Fukada;M. Kudou;K. Tsumoto;Y. Toyama;Syunsaku Kado;K. Kubota;S. Takeda
P2噬菌体病毒粒子在基板下方具有尾刺蛋白，并在感染过程中利用它们吸附到大肠杆菌的外膜上。以前的免疫电镜研究表明，尾刺是由基因V产品（gpV），然而，其膜结合活性的实验证据尚未报道。在这项研究中，我们纯化和表征重组全长gpV及其C-末端结构域。限制糜蛋白酶蛋白水解gpV产生的C-末端结构域组成的Ser 86-Leu 211。我们的实验表明，N-和C-末端结构域具有非常不同的解链温度：分别为50和74摄氏度。我们还发现gpV与E.大肠杆菌膜通过其C-末端结构域。我们得出结论，gpV的C-末端结构域是一个稳定的三聚体，并作为受体结合域的第二步中的噬菌体吸附过程。
The P2 phage virion has tail spike proteins beneath the baseplate and uses them to adsorb to the outer membrane of Escherichia coli during the infection process. Previous immunoelectron microscopic studies suggested that the tail spikes are composed of the gene V product (gpV); however, experimental evidence of its membrane binding activity has yet to be reported. In this study, we purified and characterized recombinant full-length gpV and its C-terminal domain. Limited chymotrypsin proteolysis of gpV produced a C-terminal domain composed of Ser86-Leu211. Our experiments demonstrated that the N- and C-terminal domains have very different melting temperatures: 50 and 74 degrees C, respectively. We also found that gpV binds the E. coli membrane via its C-terminal domain. We conclude that the C-terminal domain of gpV is a stable trimer and serves as the receptor-binding domain for the second step in the phage adsorption process.