Observation of the membrane binding activity and domain structure of gpV, which comprises the tail spike of bacteriophage P2.
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.
Department of Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan.
Yasuhiro
Y
Kageyama
Masanori
M
Murayama
Takashi
T
Onodera
Seiko
S
Yamada
Harumi
H
Fukada
Motonori
M
Kudou
Kouhei
K
Tsumoto
Yoshiharu
Y
Toyama
Syunsaku
S
Kado
Kenji
K
Kubota
Shigeki
S
Takeda
D017901:Bacteriophage P2 / Q000378:metabolism*; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008970:Molecular Weight; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013696:Temperature; D015678:Viral Structural Proteins / Q000737:chemistry* / Q000378:metabolism; D017376:Viral Tail Proteins / Q000737:chemistry* / Q000378:metabolism
D015678:Viral Structural Proteins; D017376:Viral Tail Proteins