title: Structural characterization of argingipain, a novel arginine-specific cysteine proteinase as a major periodontal pathogenic factor from Porphyromonas gingivalis.
abstract: Argingipain, so termed due to its peptide cleavage specificity at arginine residue, is a unique extracellular cysteine proteinase produced by the anaerobic rod Porphyromonas gingivalis, which is known as a major pathogenic factor of the progressive periodontal disease (T. Kadowaki, M. Yoneda, K. Okamoto, K. Maeda, and K. Yamamoto (1994) J. Biol. Chem. 269, 21371-21378). The catalytic specificity and functional importance of this enzyme prompted us to elucidate its structural features. A DNA fragment for argingipain was selectively amplified by polymerase chain reaction using mixed oligonucleotide primers designed from the NH2-terminal amino acid sequence of the purified enzyme. Although the extracellular mature enzyme was shown to have an apparent molecular mass of 44 kDa in gels, the nucleotide sequence of the isolated gene revealed a single gene coding for a 109-kDa precursor of argingipain. The deduced amino acid sequence exhibited no significant similarity to the sequences of representative members of the cysteine protease family. The precursor contained four functional domains: the NH2-terminal signal peptide required for the inner membrane transport; the NH2-terminal prosequence, which is assumed to stabilize the precursor structure; the proteinase domain; and the COOH-terminal hemagglutinin domain, which appears to be essential for extracellular secretion of the proteinase domain. Experiments involving the addition of the argingipain inhibitors to the culture medium of P. gingivalis suggested that the maturation of argingipain occurs intracellularly via an autocatalytic cleavage of the pro-argingipain propeptide.
authors.affiliation: Department of Pharmacology, Kyushu University Faculty of Dentistry, Fukuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Okamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Misumi
authors.forename: T
authors.initials: T
authors.lastname: Kadowaki
authors.forename: M
authors.initials: M
authors.lastname: Yoneda
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Ikehara
mesh_terms: D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001483:Base Sequence; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000187:drug effects / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010510:Periodontal Diseases / Q000209:etiology / Q000382:microbiology; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D011480:Protease Inhibitors / Q000494:pharmacology; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D017421:Sequence Analysis
chemical_list: D018829:Adhesins, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D010446:Peptide Fragments; D011480:Protease Inhibitors; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases