Processing of the AIDA-I precursor: removal of AIDAc and evidence for the outer membrane anchoring as a beta-barrel structure.
The AIDA-I adhesin known to be responsible for the diffuse adherence (DA) phenotype of the diarrhoeagenic Escherichia coli (DAEC) strain 2787 has been shown previously to be synthesized as a precursor protein and to undergo additional C-terminal processing. Here, the C-terminal processing of the AIDA-I precursor and the outer membrane topology of the cleaved C-terminal fragment, AIDAc, were investigated. By isolation of the cleaved AIDAc fragment and N-terminal sequencing, the C-terminal cleavage site was identified between Ser-846 and Ala-847 thereby indicating a molecular mass of 47.5 kDa for AIDAc. The correct processing to AIDA-I and AIDAc in OmpT, OmpP and DegP protease-deficient E. coli strains as well as in avirulent salmonellae and shigellae points to an autocatalytic cleavage mechanism. The cleaved AIDAc was localized in the outer membrane. A leader sequence-AIDAc fusion was efficiently routed to the outer membrane. Analysis by protease digestion, secondary-structure prediction and modelling, by comparison with structurally related bacterial proteins like the IgA1 protease from neisseria, the vacuolating toxin from Helicobacter pylori, and the VirG protein of Shigella flexneri, strongly indicates that AIDAc is present in the outer membrane as a beta-barrel structure.
Suhr
M
M
Institut für Infektiologie, Zentrum für Molekularbiologie, Entzündung (ZMBE), Münster, Germany.
Benz
I
I
Schmidt
M A
MA
D018830:Adhesins, Escherichia coli / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial; D015153:Blotting, Western; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000648:ultrastructure; D010450:Endopeptidases / Q000172:deficiency / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D017421:Sequence Analysis
C075559:AIDA-I protein, E coli; D018830:Adhesins, Escherichia coli; D000907:Antibodies, Bacterial; D011498:Protein Precursors; D010450:Endopeptidases