Specific interaction of fused H protein of bacteriophage phiX174 with receptor lipopolysaccharides.
The DNA fragment encoding the spike H protein of bacteriophage phiX174 was amplified by polymerase chain reaction. The fragment was sub-cloned into pQE-30 to yield pQE-H. The histidine-tagged H protein (HisH) was obtained from the cell extract of Escherichia coli M15 (pREP4) harboring pQE-H and purified by nickel chelating and anion-exchange chromatographies. HisH was shown to bind dose-dependently to the lipopolysaccharides (LPSs) isolated from phiX174-sensitive strains, E. coli C or Salmonella typhimurium TV119 (Ra mutant). In sharp contrast, HisH did not bind to the LPSs from insensitive strains, E. coli F583 (Rd2 mutant) or E. coli O111:B4 (smooth strain). Since the same selectivity was observed in the plaque counting assay for in vitro inactivation of phiX174, the spike H protein was shown to recognize receptor LPS.
Suzuki
R
R
Department of Bioscience, Mie University, Tsu, Japan.
Inagaki
M
M
Karita
S
S
Kawaura
T
T
Kato
M
M
Nishikawa
S
S
Kashimura
N
N
Morita
J
J
D010584:Bacteriophage phi X 174 / Q000378:metabolism*; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins