Open AccessOverproduced and purified receptor binding protein pb5 of bacteriophage T5 binds to the T5 receptor protein FhuA
Author(s) -
Mondigler M.,
Vögele R.T.,
Heller K.J.
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07734.x
Subject(s) - fast protein liquid chromatography , microbiology and biotechnology , biology , cytosol , biochemistry , chemistry , enzyme
A promotor‐less oad gene of bacteriophage T5, encoding the receptor binding protein pb5, was cloned into pT7‐3 under the control of phage T7 promoter Φ10. Induction with IPTG resulted in enhanced production of pb5. Upon fractionation of the producing cells, most of the overproduced pb5 was found in the membrane fraction, which was most likely due to aggregation of the protein. The minor, soluble fraction of pb5 specifically inhibited adsorption of T5 to its FhuA receptor protein. Inhibition was also seen with trace amounts of pb5, and binding of pb5 to FhuA appeared to be almost irreversible. Purification of pb5 from the cytosolic fraction was performed by FPLC using a MonoQ column. pb5, which did not bind to the matrix of the column, was obtained in almost pure form. The purified protein also inhibited T5 adsorption.