title: The PtlE protein of Bordetella pertussis has peptidoglycanase activity required for Ptl-mediated pertussis toxin secretion.
abstract: Pertussis toxin of Bordetella pertussis is secreted by a type IV secretion system comprised of the products of the nine ptl (pertussis toxin liberation) genes. These proteins are believed to form a complex spanning both the inner and outer membranes and passing through the peptidoglycan layer. Peptidoglycan acts as a barrier for transport through the periplasm of large folded molecules. Assembled pertussis toxin and the secretion component proteins PtlC through PtlH are too large to diffuse through intact peptidoglycan. Therefore, we hypothesized that the Ptl system contains a peptidoglycanase activity. The PtlE protein was found to exhibit a sequence match to the active site of glycohydrolase enzymes. An N-terminally polyhistidine-tagged PtlE fusion protein, constructed and expressed in Escherichia coli and in B. pertussis, exhibited peptidoglycanase activity on activity gels. A fusion protein with alanine substitutions at the putative active site residues (aspartic acid at position 53 and glutamic acid at position 62) lacked peptidoglycanase activity. B. pertussis strains with the amino acid substitutions were deficient for pertussis toxin secretion. Based on these results, we concluded that PtlE is a peptidoglycanase responsible for the local removal or rearrangement of the peptidoglycan layer during Ptl secretion complex assembly.
authors: Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, Cincinnati, Ohio 45267, USA.
authors: Amy A
authors: AA
authors: Rambow-Larsen
authors: Alison A
authors: AA
authors: Weiss
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D001665:Binding Sites; D001886:Bordetella pertussis / Q000201:enzymology / Q000378:metabolism*; D006224:Cricetinae; D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006639:Histidine; D009154:Mutation; D009876:Operon; D010455:Peptides; D037342:Pertussis Toxin; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D010566:Virulence Factors, Bordetella / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; C102588:PtlI protein, Bordetella pertussis; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; C033223:polyhistidine; D006639:Histidine; D037342:Pertussis Toxin; D006026:Glycoside Hydrolases