Identification and characterization of a chymotrypsin-like serine protease from periodontal pathogen, Tannerella forsythia.
Tannerella forsythia is a bacteria associated with severe periodontal disease. This study reports identification and characterization of a membrane-associated serine protease from T. forsythia. The protease was isolated from T. forsythia membrane fractions and shown to cleave both gelatin and type I collagen. The protease was able to cleave both substrates over a wide range of pH values, however optimal cleavage occurred at pH 7.5 for gelatin and 8.0 for type I collagen. The protease was also shown to cleave both gelatin and type I collagen at the average reported temperature for the gingival sulcus however it showed a lack of thermal stability with a complete loss of activity by 60 °C. When treated with protease inhibitors the enzyme's activity could only be completely inhibited by serine protease inhibitors antipain and phenylmethanesulfonyl fluoride (PMSF). Further characterization of the protease utilized serine protease synthetic peptides. The protease cleaved N-succinyl-Ala-Ala-Pro-Phe p-nitroanilide but not Nα-benzoyl-dl-arginine p-nitroanilide (BAPNA) or N-methoxysuccinyl-Ala-Ala-Pro-Val p-nitroanilide indicating that the protease is a chymotrypsin-like serine protease. Since type I collagen is a major component in the gingival tissues and periodontal ligament, identification and characterization of this enzyme provides important information regarding the role of T. forsythia in periodontal disease.
Drury University, 900 N. Benton, Springfield, MO 65802, United States.
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K
Hockensmith
Drury University, 900 N. Benton, Springfield, MO 65802, United States.
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Dillard
Drury University, 900 N. Benton, Springfield, MO 65802, United States.
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Sanders
Drury University, 900 N. Benton, Springfield, MO 65802, United States. Electronic address: bharville@drury.edu.
B A
BA
Harville
D000976:Antipain / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000032:analysis; D004795:Enzyme Stability; D005780:Gelatin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010664:Phenylmethylsulfonyl Fluoride / Q000378:metabolism; D057057:Serine Proteases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013379:Substrate Specificity; D000070436:Tannerella forsythia / Q000201:enzymology*; D013696:Temperature
D024042:Collagen Type I; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D000976:Antipain; D010664:Phenylmethylsulfonyl Fluoride; D005780:Gelatin; D057057:Serine Proteases
Gelatin; Periodontal disease; Serine protease; Tannerella forsythia; Type I collagen