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Infect Immun. Aug 1997; 65(8): 3231–3238.
PMCID: PMC175457

Cystalysin, a 46-kilodalton cysteine desulfhydrase from Treponema denticola, with hemolytic and hemoxidative activities.


A 46-kDa hemolytic protein, referred to as cystalysin, from Treponema denticola ATCC 35404 was overexpressed in Escherichia coli LC-67. Both the native and recombinant 46-kDa proteins were purified to homogeneity. Both proteins expressed identical biological and functional characteristics. In addition to its biological function of lysing erythrocytes and hemoxidizing the hemoglobin to methemoglobin, cystalysin was also capable of removing the sulfhydryl and amino groups from selected S-containing compounds (e.g., cysteine) producing H2S, NH3, and pyruvate. This cysteine desulfhydrase resulted in the following Michaelis-Menten kinetics: Km = 3.6 mM and k(cat) = 12 s(-1). Cystathionine and S-aminoethyl-L-cysteine were also substrates for the protein. Gas chromatography-mass spectrometry and high-performance liquid chromatography analysis of the end products revealed NH3, pyruvate, homocysteine (from cystathionine), and cysteamine (from S-aminoethyl-L-cysteine). The enzyme was active over a broad pH range, with highest activity at pH 7.8 to 8.0. The enzymatic activity was increased by beta-mercaptoethanol. It was not inhibited by the proteinase inhibitor TLCK (N alpha-p-tosyl-L-lysine chloromethyl ketone), pronase, or proteinase K, suggesting that the functional site was physically protected or located in a small fragment of the polypeptide. We hypothesize that cystalysin is a pyridoxal-5-phosphate-containing enzyme, with activity of an alphaC-N and betaC-S lyase (cystathionase) type. Since large amounts of H2S have been reported in deep periodontal pockets, cystalysin may also function in vivo as an important virulence molecule.

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Selected References

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  • Carlsson J, Larsen JT, Edlund MB. Peptostreptococcus micros has a uniquely high capacity to form hydrogen sulfide from glutathione. Oral Microbiol Immunol. 1993 Feb;8(1):42–45. [PubMed]
  • Chu L, Kennell W, Holt SC. Characterization of hemolysis and hemoxidation activities by Treponema denticola. Microb Pathog. 1994 Mar;16(3):183–195. [PubMed]
  • Chu L, Holt SC. Purification and characterization of a 45 kDa hemolysin from Treponema denticola ATCC 35404. Microb Pathog. 1994 Mar;16(3):197–212. [PubMed]
  • Chu L, Burgum A, Kolodrubetz D, Holt SC. The 46-kilodalton-hemolysin gene from Treponema denticola encodes a novel hemolysin homologous to aminotransferases. Infect Immun. 1995 Nov;63(11):4448–4455. [PMC free article] [PubMed]
  • Chu L, Song M, Holt SC. Effect of iron regulation on expression and hemin-binding function of outer-sheath proteins from Treponema denticola. Microb Pathog. 1994 May;16(5):321–335. [PubMed]
  • Claesson R, Edlund MB, Persson S, Carlsson J. Production of volatile sulfur compounds by various Fusobacterium species. Oral Microbiol Immunol. 1990 Jun;5(3):137–142. [PubMed]
  • Douet JP, Castroviejo M, Dodin A, Bébéar C. Purification and characterization of Kanagawa haemolysin from Vibrio parahaemolyticus. Res Microbiol. 1992 Jul-Aug;143(6):569–577. [PubMed]
  • Fiehn NE. Enzyme activities from eight small-sized oral spirochetes. Scand J Dent Res. 1986 Apr;94(2):132–140. [PubMed]
  • Grenier D. Characteristics of hemolytic and hemagglutinating activities of Treponema denticola. Oral Microbiol Immunol. 1991 Aug;6(4):246–249. [PubMed]
  • Guarneros G, Ortega MV. Cysteine desulfhydrase activities of Salmonella typhimurium and Escherichia coli. Biochim Biophys Acta. 1970 Jan 14;198(1):132–142. [PubMed]
  • Haapasalo M, Müller KH, Uitto VJ, Leung WK, McBride BC. Characterization, cloning, and binding properties of the major 53-kilodalton Treponema denticola surface antigen. Infect Immun. 1992 May;60(5):2058–2065. [PMC free article] [PubMed]
  • Holt SC, Bramanti TE. Factors in virulence expression and their role in periodontal disease pathogenesis. Crit Rev Oral Biol Med. 1991;2(2):177–281. [PubMed]
  • Horowitz A, Folke LE. Hydrogen sulfide production in the periodontal environment. J Periodontol. 1973 Jul;44(7):390–395. [PubMed]
  • Ikigai H, Akatsuka A, Tsujiyama H, Nakae T, Shimamura T. Mechanism of membrane damage by El Tor hemolysin of Vibrio cholerae O1. Infect Immun. 1996 Aug;64(8):2968–2973. [PMC free article] [PubMed]
  • Ishihara K, Kuramitsu HK. Cloning and expression of a neutral phosphatase gene from Treponema denticola. Infect Immun. 1995 Apr;63(4):1147–1152. [PMC free article] [PubMed]
  • Kennell W, Holt SC. Comparative studies of the outer membranes of Bacteroides gingivalis, strains ATCC 33277, W50, W83, 381. Oral Microbiol Immunol. 1990 Jun;5(3):121–130. [PubMed]
  • Kredich NM, Foote LJ, Keenan BS. The stoichiometry and kinetics of the inducible cysteine desulfhydrase from Salmonella typhimurium. J Biol Chem. 1973 Sep 10;248(17):6187–6196. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • LEAHY T, SMITH R. Notes on methemoglobin determination. Clin Chem. 1960 Apr;6:148–152. [PubMed]
  • Maltha JC, Mikx FH, Kuijpers FJ. Necrotizing ulcerative gingivitis in beagle dogs. III. Distribution of spirochetes in interdental gingival tissue. J Periodontal Res. 1985 Sep;20(5):522–531. [PubMed]
  • Mehta PK, Hale TI, Christen P. Evolutionary relationships among aminotransferases. Tyrosine aminotransferase, histidinol-phosphate aminotransferase, and aspartate aminotransferase are homologous proteins. Eur J Biochem. 1989 Dec 8;186(1-2):249–253. [PubMed]
  • Mehta PK, Christen P. Homology of pyridoxal-5'-phosphate-dependent aminotransferases with the cobC (cobalamin synthesis), nifS (nitrogen fixation), pabC (p-aminobenzoate synthesis) and malY (abolishing endogenous induction of the maltose system) gene products. Eur J Biochem. 1993 Jan 15;211(1-2):373–376. [PubMed]
  • Menestrina G, Moser C, Pellet S, Welch R. Pore-formation by Escherichia coli hemolysin (HlyA) and other members of the RTX toxins family. Toxicology. 1994 Feb 28;87(1-3):249–267. [PubMed]
  • Mihara J, Holt SC. Purification and characterization of fibroblast-activating factor isolated from Porphyromonas gingivalis W50. Infect Immun. 1993 Feb;61(2):588–595. [PMC free article] [PubMed]
  • Moore WE. Microbiology of periodontal disease. J Periodontal Res. 1987 Sep;22(5):335–341. [PubMed]
  • Morhart RE, Mata LJ, Sinskey AJ, Harris RS. A microbiological and biochemical study of gingival crevice debris obtained from Guatemalan Mayan Indians. J Periodontol. 1970 Nov;41(11):644–649. [PubMed]
  • Pederson ED, Miller JW, Matheson S, Simonson LG, Chadwick DE, Covill PJ, Turner DW, Lamberts BL, Morton HE. Trypsin-like activity levels of Treponema denticola and Porphyromonas gingivalis in adults with periodontitis. J Clin Periodontol. 1994 Sep;21(8):519–525. [PubMed]
  • Persson S. Hydrogen sulfide and methyl mercaptan in periodontal pockets. Oral Microbiol Immunol. 1992 Dec;7(6):378–379. [PubMed]
  • Persson S, Edlund MB, Claesson R, Carlsson J. The formation of hydrogen sulfide and methyl mercaptan by oral bacteria. Oral Microbiol Immunol. 1990 Aug;5(4):195–201. [PubMed]
  • Rams TE, Andriolo M, Jr, Feik D, Abel SN, McGivern TM, Slots J. Microbiological study of HIV-related periodontitis. J Periodontol. 1991 Jan;62(1):74–81. [PubMed]
  • Reidl J, Boos W. The malX malY operon of Escherichia coli encodes a novel enzyme II of the phosphotransferase system recognizing glucose and maltose and an enzyme abolishing the endogenous induction of the maltose system. J Bacteriol. 1991 Aug;173(15):4862–4876. [PMC free article] [PubMed]
  • Riviere GR, Weisz KS, Simonson LG, Lukehart SA. Pathogen-related spirochetes identified within gingival tissue from patients with acute necrotizing ulcerative gingivitis. Infect Immun. 1991 Aug;59(8):2653–2657. [PMC free article] [PubMed]
  • Riviere GR, Weisz KS, Adams DF, Thomas DD. Pathogen-related oral spirochetes from dental plaque are invasive. Infect Immun. 1991 Oct;59(10):3377–3380. [PMC free article] [PubMed]
  • Riviere GR, Elliot KS, Adams DF, Simonson LG, Forgas LB, Nilius AM, Lukehart SA. Relative proportions of pathogen-related oral spirochetes (PROS) and Treponema denticola in supragingival and subgingival plaque from patients with periodontitis. J Periodontol. 1992 Feb;63(2):131–136. [PubMed]
  • Riviere GR, Smith KS, Carranza N, Jr, Tzagaroulaki E, Kay SL, Dock M. Subgingival distribution of Treponema denticola, Treponema socranskii, and pathogen-related oral spirochetes: prevalence and relationship to periodontal status of sampled sites. J Periodontol. 1995 Oct;66(10):829–837. [PubMed]
  • Rizzo AA. The possible role of hydrogen sulfide in human periodontal disease. I. Hydrogen sulfide production in periodontal pockets. Periodontics. 1967 Sep-Oct;5(5):233–236. [PubMed]
  • Rosen G, Naor R, Rahamim E, Yishai R, Sela MN. Proteases of Treponema denticola outer sheath and extracellular vesicles. Infect Immun. 1995 Oct;63(10):3973–3979. [PMC free article] [PubMed]
  • Rossol I, Pühler A. The Corynebacterium glutamicum aecD gene encodes a C-S lyase with alpha, beta-elimination activity that degrades aminoethylcysteine. J Bacteriol. 1992 May;174(9):2968–2977. [PMC free article] [PubMed]
  • Saglie R, Newman MG, Carranza FA, Jr, Pattison GL. Bacterial invasion of gingiva in advanced periodontitis in humans. J Periodontol. 1982 Apr;53(4):217–222. [PubMed]
  • Saglie R, Newman MG, Carranza FA., Jr A scanning electron microscopic study of leukocytes and their interaction with bacteria in human periodontitis. J Periodontol. 1982 Dec;53(12):752–761. [PubMed]
  • Scott D, Siboo IR, Chan EC, Klitorinos A, Siboo R. Binding of hemin and congo red by oral hemolytic spirochetes. Oral Microbiol Immunol. 1993 Aug;8(4):245–250. [PubMed]
  • SIEGEL LM. A DIRECT MICRODETERMINATION FOR SULFIDE. Anal Biochem. 1965 Apr;11:126–132. [PubMed]
  • Simonson LG, McMahon KT, Childers DW, Morton HE. Bacterial synergy of Treponema denticola and Porphyromonas gingivalis in a multinational population. Oral Microbiol Immunol. 1992 Apr;7(2):111–112. [PubMed]
  • Simonson LG, Goodman CH, Bial JJ, Morton HE. Quantitative relationship of Treponema denticola to severity of periodontal disease. Infect Immun. 1988 Apr;56(4):726–728. [PMC free article] [PubMed]
  • Simonson LG, Goodman CH, Morton HE. Quantitative immunoassay of Treponema denticola serovar C in adult periodontitis. J Clin Microbiol. 1990 Jul;28(7):1493–1496. [PMC free article] [PubMed]
  • Sung MH, Tanizawa K, Tanaka H, Kuramitsu S, Kagamiyama H, Hirotsu K, Okamoto A, Higuchi T, Soda K. Thermostable aspartate aminotransferase from a thermophilic Bacillus species. Gene cloning, sequence determination, and preliminary x-ray characterization. J Biol Chem. 1991 Feb 5;266(4):2567–2572. [PubMed]
  • Syed SA, Mäkinen KK, Mäkinen PL, Chen CY, Muhammad Z. Proteolytic and oxidoreductase activity of Treponema denticola ATCC 35405 grown in an aerobic and anaerobic gaseous environment. Res Microbiol. 1993 May;144(4):317–326. [PubMed]
  • Umemoto T, Nakazawa F, Hoshino E, Okada K, Fukunaga M, Namikawa I. Treponema medium sp. nov., isolated from human subgingival dental plaque. Int J Syst Bacteriol. 1997 Jan;47(1):67–72. [PubMed]
  • Walker B, Bayley H. Key residues for membrane binding, oligomerization, and pore forming activity of staphylococcal alpha-hemolysin identified by cysteine scanning mutagenesis and targeted chemical modification. J Biol Chem. 1995 Sep 29;270(39):23065–23071. [PubMed]
  • Weinberg A, Holt SC. Interaction of Treponema denticola TD-4, GM-1, and MS25 with human gingival fibroblasts. Infect Immun. 1990 Jun;58(6):1720–1729. [PMC free article] [PubMed]
  • Weinberg A, Holt SC. Chemical and biological activities of a 64-kilodalton outer sheath protein from Treponema denticola strains. J Bacteriol. 1991 Nov;173(21):6935–6947. [PMC free article] [PubMed]
  • Welch RA. Pore-forming cytolysins of gram-negative bacteria. Mol Microbiol. 1991 Mar;5(3):521–528. [PubMed]
  • Wilmsen HU, Pattus F, Buckley JT. Aerolysin, a hemolysin from Aeromonas hydrophila, forms voltage-gated channels in planar lipid bilayers. J Membr Biol. 1990 Apr;115(1):71–81. [PubMed]
  • Wyss C, Choi BK, Schüpbach P, Guggenheim B, Göbel UB. Treponema maltophilum sp. nov., a small oral spirochete isolated from human periodontal lesions. Int J Syst Bacteriol. 1996 Jul;46(3):745–752. [PubMed]
  • Zdych E, Peist R, Reidl J, Boos W. MalY of Escherichia coli is an enzyme with the activity of a beta C-S lyase (cystathionase). J Bacteriol. 1995 Sep;177(17):5035–5039. [PMC free article] [PubMed]
  • Zheng L, White RH, Cash VL, Jack RF, Dean DR. Cysteine desulfurase activity indicates a role for NIFS in metallocluster biosynthesis. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2754–2758. [PMC free article] [PubMed]

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