Purification and Characterization of Two Listeria Ivanovii Cytolysins, a Sphingomyelinase C and a Thiol-activated Toxin (ivanolysin O)

Overview

Journal Infect Immun

Publisher American Society for Microbiology

Specialty Allergy & Immunology

Date 1989 Dec 1

PMID 2553614

Citations 11

Authors

J A Vazquez-Boland

L Dominguez

E F Rodriguez-Ferri

G Suarez

Affiliations

Soon will be listed here.

Abstract

The strong bizonal hemolysis on blood agar and the positive CAMP reaction with Rhodococcus equi denotes the production of two different cytolytic factors by Listeria ivanovii. One was characterized as a thiol-activated (SH) cytolysin of 61 kilodaltons and was termed ivanolysin O (ILO) since data suggested that it is different from listeriolysin O, the SH-cytolysin produced by Listeria monocytogenes. The other is a 27-kilodalton hemolytic sphingomyelinase C that was found to be the cytolytic factor responsible for the halo of incomplete hemolysis synergistically enhanced by R. equi exosubstances. When thiol-disulfide exchange affinity chromatography and gel filtration were applied to the purification of ILO from concentrated L. ivanovii culture supernatants, the copurification of the two cytolysins was observed. This phenomenon seems to be due to the formation of intermolecular disulfide bonds between ILO and the sphingomyelinase, since the latter was found to contain free SH groups, not essential for the activity. These SH groups could react with the single cysteine residue characteristically present in the SH-cytolysins, forming a dimeric cytolytic complex. The purification of ILO was achieved by a further gel filtration with a reducing agent (dithiothreitol) in the eluent. A method for the purification of the sphingomyelinase based on selective sequestration of ILO from the L. ivanovii concentrated culture supernatant by the SH cytolysin target molecule cholesterol and thiol-disulfide affinity chromatography is described.

Citing Articles

Differences in gamma interferon production induced by listeriolysin O and ivanolysin O result in different levels of protective immunity in mice infected with Listeria monocytogenes and Listeria ivanovii.

Kimoto T, Kawamura I, Kohda C, Nomura T, Tsuchiya K, Ito Y Infect Immun. 2003; 71(5):2447-54.

PMID: 12704115 PMC: 153848. DOI: 10.1128/IAI.71.5.2447-2454.2003.

A DNA delivery system containing listeriolysin O results in enhanced hepatocyte-directed gene expression.

Walton C, Wu C, Wu G World J Gastroenterol. 2002; 5(6):465-469.

PMID: 11819493 PMC: 4688787. DOI: 10.3748/wjg.v5.i6.465.

Listeria pathogenesis and molecular virulence determinants.

Vazquez-Boland J, Kuhn M, Berche P, Chakraborty T, Dominguez-Bernal G, Goebel W Clin Microbiol Rev. 2001; 14(3):584-640.

PMID: 11432815 PMC: 88991. DOI: 10.1128/CMR.14.3.584-640.2001.

Production of monoclonal antibodies to Listeria monocytogenes and their application to determine the virulence of isolates from channel catfish.

Erdenlig S, Ainsworth A, Austin F Appl Environ Microbiol. 1999; 65(7):2827-32.

PMID: 10388671 PMC: 91424. DOI: 10.1128/AEM.65.7.2827-2832.1999.

Influence of environmental parameters on phosphatidylcholine phospholipase C production in Listeria monocytogenes: a convenient method to differentiate L. monocytogenes from other Listeria species.

Coffey A, Rombouts F, Abee T Appl Environ Microbiol. 1996; 62(4):1252-6.

PMID: 8919785 PMC: 167890. DOI: 10.1128/aem.62.4.1252-1256.1996.

References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Kreft J, Funke D, Haas A, Lottspeich F, Goebel W . Production, purification and characterization of hemolysins from Listeria ivanovii and Listeria monocytogenes Sv4b. FEMS Microbiol Lett. 1989; 48(2):197-202. DOI: 10.1111/j.1574-6968.1989.tb03298.x. View

Cooper R, DENNIS S . Further characterization of Listeria monocytogenes serotype 5. Can J Microbiol. 1978; 24(5):598-9. DOI: 10.1139/m78-097. View

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

Johnson M, Geoffroy C, ALOUF J . Binding of cholesterol by sulfhydryl-activated cytolysins. Infect Immun. 1980; 27(1):97-101. PMC: 550729. DOI: 10.1128/iai.27.1.97-101.1980. View

BERNHEIMER A, Linder R, Avigad L . Stepwise degradation of membrane sphingomyelin by corynebacterial phospholipases. Infect Immun. 1980; 29(1):123-31. PMC: 551084. DOI: 10.1128/iai.29.1.123-131.1980. View

SKALKA B, Smola J, ELISCHEROVA K . Routine test for in vitro differentiation of pathogenic and apathogenic Listeria monocytogenes strains. J Clin Microbiol. 1982; 15(3):503-7. PMC: 272125. DOI: 10.1128/jcm.15.3.503-507.1982. View

Tomita M, Taguchi R, Ikezawa H . Molecular properties and kinetic studies on sphingomyelinase of Bacillus cereus. Biochim Biophys Acta. 1982; 704(1):90-9. DOI: 10.1016/0167-4838(82)90135-2. View

BERNHEIMER A, Avigad L . Mechanism of hemolysis by Renalin, a CAMP-like protein from Corynebacterium renale. Infect Immun. 1982; 36(3):1253-6. PMC: 551466. DOI: 10.1128/iai.36.3.1253-1256.1982. View

10.

SEELIGER H, Schrettenbrunner A, Pongratz G, Hof H . [Special position of strongly haemolytic strains of the genus Listeria]. Zentralbl Bakteriol Mikrobiol Hyg A Med Mikrobiol Infekt Parasitol. 1982; 252(2):176-90. View

11.

Rocourt J, Alonso J, SEELIGER H . [Comparative virulence of the 5 genomic groups of Listeria monocytogenes (sensu lato)]. Ann Microbiol (Paris). 1983; 134A(3):359-64. View

12.

Linder R . Alteration of mammalian membranes by the cooperative and antagonistic actions of bacterial proteins. Biochim Biophys Acta. 1984; 779(4):423-35. DOI: 10.1016/0304-4157(84)90019-4. View

13.

Randall L, Hardy S . Export of protein in bacteria. Microbiol Rev. 1984; 48(4):290-8. PMC: 373220. DOI: 10.1128/mr.48.4.290-298.1984. View

14.

Parrisius J, Bhakdi S, Roth M, Tranum-Jensen J, Goebel W, SEELIGER H . Production of listeriolysin by beta-hemolytic strains of Listeria monocytogenes. Infect Immun. 1986; 51(1):314-9. PMC: 261104. DOI: 10.1128/iai.51.1.314-319.1986. View

15.

Rocourt J, Catimel B . [Biochemical characterization of species in the genus Listeria]. Zentralbl Bakteriol Mikrobiol Hyg A. 1985; 260(2):221-31. View

16.

Gaillard J, Berche P, Sansonetti P . Transposon mutagenesis as a tool to study the role of hemolysin in the virulence of Listeria monocytogenes. Infect Immun. 1986; 52(1):50-5. PMC: 262196. DOI: 10.1128/iai.52.1.50-55.1986. View

17.

BERNHEIMER A, Rudy B . Interactions between membranes and cytolytic peptides. Biochim Biophys Acta. 1986; 864(1):123-41. DOI: 10.1016/0304-4157(86)90018-3. View

18.

Dominguez Rodriguez L, Vazquez Boland J, Fernandez Garayzabal J, Echalecu Tranchant P, Gomez-Lucia E, Rodriguez Ferri E . Microplate technique to determine hemolytic activity for routine typing of Listeria strains. J Clin Microbiol. 1986; 24(1):99-103. PMC: 268840. DOI: 10.1128/jcm.24.1.99-103.1986. View

19.

Kathariou S, Metz P, Hof H, Goebel W . Tn916-induced mutations in the hemolysin determinant affecting virulence of Listeria monocytogenes. J Bacteriol. 1987; 169(3):1291-7. PMC: 211933. DOI: 10.1128/jb.169.3.1291-1297.1987. View

20.

Walker J, Allen R, Falmagne P, Johnson M, Boulnois G . Molecular cloning, characterization, and complete nucleotide sequence of the gene for pneumolysin, the sulfhydryl-activated toxin of Streptococcus pneumoniae. Infect Immun. 1987; 55(5):1184-9. PMC: 260488. DOI: 10.1128/iai.55.5.1184-1189.1987. View