title: Functional analysis of the thioredoxin domain in Porphyromonas gingivalis HBP35.
abstract: Periodontitis is one of the most common oral diseases in humans. This caused by infection by the oral bacterium Porphyromonas gingivalis. Our strategy to prevent this infection is to establish a passive immunization system in which endogenous antibodies can be applied directly to neutralize virulent factors associated with this bacterium. We focused our attention on the P. gingivalis 35 kDa surface protein, or HBP35, since this protein is involved not only in the coaggregation with oral miroflora but also in hemin binding. In addition, nucleotide sequencing of the gene, hbp35, coding for this protein revealed the presence of a catalytic center for thioredoxin, and we further attempted to characterized the protein by amino acid substitution. A total of four Cys residues were substituted for Ser residues by combining the simple method for site-directed mutagenesis and the heterodimer system, an approach designed to construct chimeric plasmids readily. Native and mutagenized hbp35 were introduced into the Eschericha coli dsbA mutant strain, JCB 572, defective in both alkaline phosphatase and motile activities due to inefficient disulfide bond formation. Transformant harboring the native hbp35 could complement the dsbA mutation, suggesting a role of disulfide bond formation of this protein in P. gingivalis cells. Possible roles of the Cys residues in complementation are discussed.
authors.affiliation: Department of Chemistry, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, Japan. shiroza.teruaki@nihon-u.ac.jp
authors.forename: Teruaki
authors.initials: T
authors.lastname: Shiroza
authors.forename: Soichiro
authors.initials: S
authors.lastname: Okano
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Shibata
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Hayakawa
authors.forename: Kenichi
authors.initials: K
authors.lastname: Fujita
authors.forename: Kosuke
authors.initials: K
authors.lastname: Yamaguchi
authors.forename: Yoshimitsu
authors.initials: Y
authors.lastname: Abiko
mesh_terms: D019943:Amino Acid Substitution; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D020134:Catalytic Domain; D004220:Disulfides; D004926:Escherichia coli; D005816:Genetic Complementation Test; D000081062:Heme-Binding Proteins; D006420:Hemeproteins / Q000235:genetics* / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D016966:Porphyromonas gingivalis / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D013879:Thioredoxins / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D004220:Disulfides; D000081062:Heme-Binding Proteins; D006420:Hemeproteins; D013879:Thioredoxins