Need for TolC, an Escherichia coli outer membrane protein, in the secretion of heat-stable enterotoxin I across the outer membrane.
Escherichia coli heat-stable enterotoxin Ip (STIp) is a typical extracellular toxin consisting of 18 amino acid residues synthesized as a precursor of pre (amino acid residues 1 to 19), pro (amino acid residues 20 to 54), and mature (amino acid residues 55 to 72) regions. STIp synthesized in the cytoplasm must cross the inner and outer membranes to migrate into the extracellular environment. Previous studies showed that the precursor translocates across the inner membrane utilizing the general export pathway consisting of Sec proteins. However, it remains unclear how it crosses the outer membrane. In this study, we examined the effects of mutation of the tolC gene which encodes an E. coli outer membrane protein, TolC, on the release of STIp into the extracellular environment. The mutation reduced the amount of STIp released into culture supernatant and increased the amount of STIp accumulated in the periplasm. This indicates that TolC mediates the translocation of STIp across the outer membrane. The inability to transfer STIp in the periplasm into the culture supernatant was restored by introduction of the tolC gene into the mutant cells. In the mouse intestinal loop assay, living cells of the mutants did not show a positive response, but wild-type cells did. These results showed that TolC is involved in the translocation of STIp across the outer membrane.
Yamanaka
H
H
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima, Yamashiro, Tokushima, 770-8514, Japan.
Nomura
T
T
Fujii
Y
Y
Okamoto
K
K
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000502:physiology*; D001427:Bacterial Toxins / Q000378:metabolism* / Q000633:toxicity; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism*; D003001:Cloning, Molecular; D004768:Enterotoxins / Q000378:metabolism* / Q000633:toxicity; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006801:Humans; D026901:Membrane Transport Proteins; D051379:Mice; D016254:Mutagenesis, Insertional; D019897:Periplasm; D010957:Plasmids / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C034151:heat stable toxin (E coli); C073126:tolC protein, E coli