Drug – bio-affecting and body treating compositions – Enzyme or coenzyme containing – Hydrolases
Patent
1995-10-10
1998-01-13
Patterson, Jr., Charles L.
Drug, bio-affecting and body treating compositions
Enzyme or coenzyme containing
Hydrolases
435 21, 435 23, 435220, 53038826, 5303894, A61K 3848, C12N 952
Patent
active
057076200
ABSTRACT:
Provide herein is a substantially pure Lys-gingipain complex preparation, Lys-gingipain being characterized as having an apparent molecular mass of 105 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, where sample is prepared without boiling, said Lys-gingipain having amidolytic and proteolytic activity for cleavage after lysine residues and having no amidolytic and/or proteolytic activity for cleavage after arginine residues, wherein the amidolytic and/or proteolytic activity is inhibited by TLCK, cysteine protease group-specific inhibitors including iodoacetamide and iodoacetic acid, wherein the amidolytic and/or proteolytic activity of said Lys-gingipain is not sensitive to inhibition by leupeptin, antipain, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, serine protease group-specific inhibitors including diisopropylfluorophosphate and phenylmethyl sulfonylfluoride, and antibodies specific for the Lys-gingipain protein complex and its catalytic component, methods for preparation. As specifically exemplified, a Lys-gingipain protein complex is purified from Porphyromonas gingivalis H66, and the 60 kDa catalytic component of the Lys-gingipain protein complex has an amino acid sequence as given in SEQ ID NO:14 from amino acid 1 through amino acid 509. Also provided are nucleic acid sequences encoding this catalytic protein. The nucleotide coding sequence of the 60 kDa catalytic component of the Lys-gingipain protein complex is given in SEQ ID NO:13, from nucleotide 1336 through nucleotide 2863. The Lys-gingipain complex also comprises a hemagglutinin component identified by an N-terminal amino acid sequence as given in SEQ ID NO:14, amino acids 510-714.
REFERENCES:
Scott et al. (1993), "Purification and Characterization of a Potent 70-kDa Thiol Lysyl-proteinase (Lys-gingivain) from Porphyromonas gingivalis That Cleaves Kininogens and Fibrinogen", J. Biol. Chem. 268: 7935-7942.
Travis et al. (1993) "Activation of the Complement and Kinin Pathways by Non-Mammalian Proteinases," Abstract, Kinin 93 Brazil, 21.16.
Chen et al. (1991) "Stimulation of Proteinase and Amidase Activities in Porphyromonas (Bacteroides) gingivalis by Amino Acids and Dipeptides,"Infect. Immun. 59: 2846-2850.
Birkedal-Hansen et al. (1988) "Characterization of Collagenolytic Activity from Strains of Bacteroides gingivalis," J. Periodontal Res. 23: 258-264.
Grenier et al. (1989) "Characterization of Sodium Dodecyl Sulfate-Stable Bacteroides gingivalis Proteases by Polyacrylamide Gel Electrophoresis," Infect. Immun. 57: 95-99.
Ono et al. (1987) "Purification and Characterization of a Thiol-protease from Bacteroides gingivalis Strain 381"; Oral Micr. Immun. 2:77-81.
Madden et al. (1992), "Expression of Porphyromonas gingivalis proteolytic activity in Escherichia coli", Oral Micro. Imm. 7: 349-356.
Uitto V.J. (1987) "Human Gingival Proteases. 1: Extraction and Preliminary Characterization of Trypsin-like and Elastase-like Enzymes"; J. Periodontal Res. 22:58-63.
Sorsa et al. (1987) "A Trypsin-like Protease from Bacteroides Gingivalis; Partial Purification and Characterization"; J. Periodont Res. 22:375-380.
Hinode et al. (1992) "Genaration of Plasma Kinin by Three Types of Protease Isolated from Porphyromonas gingivalis 381," Archs Oral Biol. 10 (37):859-861.
Bourgeau et al. (1992) "Cloning, expression and sequencing of a protease gene (tpr) from Porphyromonas gingivalis W83 in Escherichia coli," Infect. Immun. 60:3186-3192.
Scott, C.F., et al. (1993) J Biol Chem. 268(11), 7935-7942.
Fujimura, S., et al. (1993) FEMS Microbiol. Letts. 113, 113-138.
Pike, R., et al. (1994) J. Biol. Chem. 269(1), 408-411.
Wingrove, J.A. (1992) J. Biol. Chem. 267(26), 18902-18907.
Chen, Z., et al. (1992) J. Biol. Chem. 267(26) 18896-18901.
Pike Robert Neil
Potempa Jan Stanislaw
Travis James
Patterson Jr. Charles L.
University of Georgia Research Foundation Inc.
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