Functional activities of the Tsh protein from avian pathogenic Escherichia coli (APEC) strains.
The temperature-sensitive hemagglutinin (Tsh) expressed by strains of avian pathogenic Escherichia (E.) coli (APEC) has both agglutinin and protease activities. Tsh is synthesized as a 140 kDa precursor protein, whose processing results in a 106 kDa passenger domain (Tsh(s)) and a 33 kDa β-domain (Tsh(β)). In this study, both recombinant Tsh (rTsh) and supernatants from APEC, which contain Tsh(s) (106 kDa), caused proteolysis of chicken tracheal mucin. Both rTsh (140 kDa) and pellets from wild-type APEC, which contain Tsh(β) (33 kDa), agglutinated chicken erythrocytes. On Western blots, the anti-rTsh antibody recognized the rTsh and 106 kDa proteins in recombinant E. coli BL21/pET 101-Tsh and in the supernatants from APEC grown at either 37°C or 42°C. Anti-rTsh also recognized a 33 kDa protein in the pellets from APEC13 cultures grown in either Luria-Bertani agar, colonization factor antigen agar, or mucin agar at either 26°C, 37°C, or 42°C, and in the extracts of outer membrane proteins of APEC. The 106 kDa protein was more evident when the bacteria were grown at 37°C in mucin agar, and it was not detected when the bacteria were grown at 26°C in any of the culture media used in this study. Chicken anti-Tsh serum inhibited hemagglutinating and mucinolytic activities of strain APEC13 and recombinant E. coli BL21/pET101-Tsh. This work suggests that the mucinolytic activity of Tsh might be important for the colonization of the avian tracheal mucous environment by APEC.
Departamento de Microbiologia-CCB, Campus Universitário, Universidade Estadual de Londrina, Caixa postal 6001, 86051-970 Londrina, Paraná, Brazil.
Renata K
RK
Kobayashi
Luis Carlos
LC
Gaziri
Marilda C
MC
Vidotto
D018830:Adhesins, Escherichia coli / Q000378:metabolism*; D001938:Brazil; D004926:Escherichia coli / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006384:Hemagglutination; D009077:Mucins / Q000378:metabolism; D021381:Protein Transport; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D018830:Adhesins, Escherichia coli; D009077:Mucins; D011994:Recombinant Proteins; C112027:tsh protein, E coli