Abstract
Aim:
To identify and classify all potential hemolysin candidates of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai.
Methods:
All of the potential hemolysin encoding genes were characterized in silico. These genes were cloned and expressed in Escherichia coli. The hemolytic activities of the expressed proteins were assayed observing the hemolysis on sheep blood agar plates. Sphingomyelinase activities of the hemolysin candidates were measured by thin-layer chromatography (TLC) and HPLC for sphingomyelin-hydrolysis. Expression and secretion of the hemolysins in L interrogans were studied by reverse transcription polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assays.
Results and Conclusion:
The hemolytic activities of hemolysin candidates (LA0327, LA0378, LA1027, LA1029, LA1650, LA3050, LA3 937, LA4004) from L interrogans strain Lai were confirmed. They were further divided into two groups, sphingomyelinase hemolysins and non-sphingomyelinase hemolysins, based on their ability to hydrolyze sphingomyelin. Most of these hemolysins were actually expressed in living L interrogans and some of them were secreted into the environment. This study establishes an essential and complete basis for further studying the contribution of hemolysins to the patho-genesis of L interrogans.
Similar content being viewed by others
Article PDF
References
Faine S . Leptospira and leptospirosis. New York: CRC Press; 1994.
Kmety E, Dikken H . Classification of the species Leptospira interrogans and history of its serovars. Groningen: University Press; 1993.
Ramadass P, Jarvis BD, Corner RJ, Penny D, Marshall RB . Genetic characterization of pathogenic Leptospira species by DNA hybridization. Int J Syst Bacteriol 1992; 42: 215–9.
Ozdemir V, Erol E . Leptospirosis in Turkey. Vet Rec 2002; 150: 248–9.
Lee SH, Kim S, Park SC, Kim MJ . Cytotoxic activities of Leptospira interrogans hemolysin SphH as a pore-forming protein on mammalian cells. Infect Immun 2002; 70: 315–22.
Sakurai J, Honda T, Jinguji Y, Arita M, Miwatani T . Cytotoxic effect of the thermostable direct hemolysin produced by Vibrio parahaemolyticus on FL cells. Infect Immun 1976; 13: 876–83.
Lee SH, Kim KA, Park YG, Seong IW, Kim MJ, Lee YJ . Identification and partial characterization of a novel hemolysin from Leptospira interrogans serovarlai. Gene 2000; 254: 19–28.
Ren SX, Fu G, Jiang XG, Zeng R, Miao YG, Xu H, et al. Unique physiological and pathogenic features of Leptospira interrogans revealed by whole genome sequencing. Nature 2003; 422: 888–93.
SWISS-PROT/TrEMBL databases. Europe: Swiss Institute of Bioinformatics. c2004. Available from: http://www.expasy.org/sprot and http://www.expasy.org/TrEMBL.
Pfam database. St Louis: Washington University in St Louis. c2004. Available from: http://pfam.wustl.edu/hmmsearch.shtml.
PROSITE database. Europe: Swiss Institute of Bioinformatics. c2004. Available from: http://us.expasy.org/prosite.
ProDom database. Europe: Laboratoire des Interactions Plantes Microorganismes. c2004. Available from: http://prodes.toulouse.inra.fr/prodom/current/html/home.php.
Jpred2 database. Scotland: University of Dundee. c2004. Available from: http://www.compbio.dundee.ac.uk/~www-jpred/submit.html.
Alexander AD, Smith OH, Hiatt CW, Gleiser C A . Presence of hemolysin in cultures of pathogenic leptospires. Proc Soc Exp Biol Med 1956; 91: 205–11.
Bauer DC, Morse EV . Variation and hemolysin production in relation to virulence of Leptospira pomona. Proc Soc Exp Biol Med 1958; 98: 505–8.
Cloning of a hemolysin gene from Leptospira interrogans serovar hardjo. Infect Immun 1989; 57: 2588–90.
Fussle R, Bhakdi S, Sziegoleit A, Tranum-Jensen J, Kranz T, Wellensiek HJ . On the mechanism of membrane damage by Staphylococcus aureus alpha-toxin. J Cell Biol 1981; 91: 83–94.
Bhakdi S, Tranum-Jensen J . Damage to cell membranes by pore-forming bacterial cytolysins. Prog Allergy 1988; 40: 1–43.
Author information
Authors and Affiliations
Corresponding author
Additional information
Project supported by the National Natural Science Foundation of China (No 30370071) and the National High Technology Research and Development Program of China (863 Program, No 2003, AA223030).
Rights and permissions
About this article
Cite this article
Zhang, Yx., Geng, Y., Bi, B. et al. Identification and classification of all potential hemolysin encoding genes and their products from Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai. Acta Pharmacol Sin 26, 453–461 (2005). https://doi.org/10.1111/j.1745-7254.2005.00075.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2005.00075.x
Keywords
This article is cited by
-
Evaluation of the Expression and Protective Potential of Leptospiral Sphingomyelinases
Current Microbiology (2010)
-
Characterization of a Hemolysin Gene ytjA from Bacillus subtilis
Current Microbiology (2009)
-
Genetic diversity among major endemic strains of Leptospira interrogans in China
BMC Genomics (2007)
-
Comparison of invasion of fibroblasts and macrophages by high- and low-virulence Leptospira strains: colonization of the host-cell nucleus and induction of necrosis by the virulent strain
Archives of Microbiology (2007)
-
Thrombocytopenia in the experimental leptospirosis of guinea pig is not related to disseminated intravascular coagulation
BMC Infectious Diseases (2006)