title: Isolation and characterization of the Porphyromonas gingivalis prtT gene, coding for protease activity.
abstract: The prtT gene, coding for trypsinlike proteolytic activity, has been isolated from Porphyromonas gingivalis ATCC 53977. This gene is present immediately downstream from the sod gene on a 5.9-kb DNA fragment from the organism isolated in Escherichia coli. The complete nucleotide sequence of the gene was determined, and the deduced amino acid sequence of the enzyme corresponds to a 53.9-kDa protein with an estimated pI of 11.85. Gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis zymography also indicated a similar molecular size for the protease. The enzyme was purified to near homogeneity following anion-exchange and gel-filtration chromatography. The purified enzyme also exhibited a single protein species with a size of approximately 53 kDa. Enzyme activity was strongly dependent upon the presence of reducing agents (dithiothreitol, cysteine, and 2-mercaptoethanol) and was also stimulated in the presence of calcium ions. A comparison of the properties of the prtT gene product with comparable parameters of proteases previously purified from different strains of P. gingivalis suggested that the cloned protease represents a previously uncharacterized enzyme.
authors.affiliation: Department of Pediatric Dentistry, University of Texas Health Science Center, San Antonio 78284-7888.
authors.forename: J
authors.initials: J
authors.lastname: Otogoto
authors.forename: H K
authors.initials: HK
authors.lastname: Kuramitsu
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D012150:Polymorphism, Restriction Fragment Length; D016966:Porphyromonas gingivalis / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014357:Trypsin / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D014357:Trypsin; D003546:Cysteine Endopeptidases; C078912:prtT protein, Porphyromonas gingivalis