Purification and characterization of a protease from Bacteroides gingivalis 381.
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An intracellular membrane-free, trypsinlike protease was isolated from cells of Bacteroides gingivalis 381 by ultrasonic treatment and was purified about 250-fold with a recovery of 2% by sequential procedures.Abstract:
An intracellular membrane-free, trypsinlike protease was isolated from cells of Bacteroides gingivalis 381. The protease was extracted from the cells by ultrasonic treatment and was purified about 250-fold with a recovery of 2% by sequential procedures. The properties of the protease were as follows: its optimal pH was 8.5; its activity was almost completely lost on incubation at 50 degrees C for 15 min; its activity was inhibited by diisopropylfluorophosphate, p-toluenesulfonyl-L-lysine chloromethyl ketone hydrochloride, leupeptin, Mn2+, Cu2+, and Zn2+; it hydrolyzed casein, azocasein, N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), bovine serum albumin, azocoll, and gelatin, but not N-alpha-benzoyl-DL-lysine-p-nitroanilide or human serum immunoglobulin A; its molecular weight was estimated as 45,000 by gel filtration and 50,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; and its Km values for azocasein and BAPNA were 1.11% and 0.19 mM, respectively.read more
Citations
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Virulence factors of Porphyromonas gingivalis.
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Biology of asaccharolytic black-pigmented Bacteroides species.
D Mayrand,S C Holt +1 more
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Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization, and evidence for the existence of complexes with hemagglutinins.
TL;DR: Two proteinases with arginine and lysine specificity were isolated from a high molecular mass fraction of the P. gingivalis culture fluid and Lys-gingipain was found to be a cysteine proteinase with optimal activity and stability at pH 8.0-8.5 and was extensively characterized in terms of its specificity and activation characteristics.
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Factors in virulence expression and their role in periodontal disease pathogenesis.
TL;DR: The classic progression of the development of periodontitis with its associated formation of an inflammatory lesion is characterized by a highly reproducible microbiological progression of a Gram-positive microbiota to a highly pathogenic Gram-negative one.
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The multiple forms of trypsin-like activity present in various strains of Porphyromonas gingivalis are due to the presence of either Arg-gingipain or Lys-gingipain.
TL;DR: Western blot (immunoblot) analysis using antibodies produced against RGP and the N-terminal peptides of RGP or the catalytic subunit of KGP indicated that these enzymes are synthesized by the strains studied and exist as multiple molecular mass species.
References
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The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis
Klaus Weber,Mary Osborn +1 more
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Estimation of the molecular weights of proteins by Sephadex gel-filtration.
TL;DR: The results are similar to those of previous studies, where the objective was to establish a cause-and-effect relationship, rather than a straightforward relationship between the number of cells and the content of the molecule.
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The preparation and properties of two new chromogenic substrates of trypsin.
TL;DR: Preliminary studies indicate that benzoyl dl -arginine p -nitroanilide hydrochloride is also hydrolyzed by papain, and that of l -LPA is in a more alkaline region than normally found for trypsin substrates.