title: The Clostridium ramosum IgA proteinase represents a novel type of metalloendopeptidase.
abstract: Clostridium ramosum is part of the normal flora in the human intestine. Some strains produce an IgA proteinase that specifically cleaves human IgA1 and the IgA2m(1) allotype. This prolylendopeptidase was purified from a broth culture supernatant, and N-terminal sequences of the native protein and tryptic fragments thereof were determined. A fragment of the iga gene encoding the IgA proteinase was isolated using degenerate primers in PCR, and the complete gene was obtained by inverse PCR. The identity of the iga gene was confirmed by heterologous expression in Escherichia coli. The deduced amino acid sequence indicated a signal peptide of 30 residues and a secreted proteinase of 133,828 Da. A typical Gram-positive cell wall anchor motif was identified in the C terminus. The presence of a putative zinc-binding motif His-Glu-Phe-Gly-His together with inhibition studies indicate that the proteinase belongs to the zinc-dependent metalloproteinases. However, the sequence of the C. ramosum IgA proteinase shows no overall similarity to other proteins except for significant identity around the zinc-binding motif with family M6 of metalloendopeptidases, and the unique sequence of the IgA proteinase in this area presumably establishes a new subfamily. The GC percentage of the iga gene is significantly higher than that for the entire genome of C. ramosum, suggesting that the gene was acquired recently in evolution.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus C DK-8000, Denmark.
authors.forename: Klaudia
authors.initials: K
authors.lastname: Kosowska
authors.forename: Jesper
authors.initials: J
authors.lastname: Reinholdt
authors.forename: Lone Kjar
authors.initials: LK
authors.lastname: Rasmussen
authors.forename: Artur
authors.initials: A
authors.lastname: Sabat
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
authors.forename: Mogens
authors.initials: M
authors.lastname: Kilian
authors.forename: Knud
authors.initials: K
authors.lastname: Poulsen
mesh_terms: D000409:Alanine / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002384:Catalysis; D002473:Cell Wall / Q000378:metabolism; D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D019143:Evolution, Molecular; D005340:Fibrinogen / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D007070:Immunoglobulin A / Q000378:metabolism; D008666:Metalloendopeptidases / Q000737:chemistry*; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000737:chemistry*; D013997:Time Factors; D015032:Zinc / Q000378:metabolism
chemical_list: D007070:Immunoglobulin A; D020033:Protein Isoforms; D011994:Recombinant Proteins; D005780:Gelatin; D005340:Fibrinogen; D009113:Muramidase; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase; D008666:Metalloendopeptidases; D015032:Zinc; D000409:Alanine