Purification of the cellular receptor of an oyster juice borne coliphage OJ367 from the outer membrane of Escherichia coli host

Abstract

The cellular receptor of an oyster juice borne phage OJ367 was found in the outer membrane (OM) of its host Escherichia coli. The total cell envelope (TCE) was fractionated by differential extraction and it was found that the OM possesses phage neutralization ability. The OM was purified by diethylaminoethyl cellulose column chromatography to screen for the phage recognition moiety. A homogeneous, 39‐kDa OM protein (Omp) with receptor activity was eluted. It was a peptidoglycan (PG)‐associated protein which showed trypsin resistance. Lipopolysaccharide (LPS) had no effect on its receptor activity either when coexisting in the column fractions or when the isolated LPS was mixed with the Omp. Mutants resistant to lysis by phage OJ367 were isolated. The amount of the 39‐kDa and another PG‐associated 37‐kDa protein decreased significantly in the TCE of the mutant. The 39‐kDa Omp may serve as the cellular receptor for adherence of the coliphage, whereas the 37‐kDa protein is also involved in the infection mechanism.