SYSTEM OF ACTIVATION OF PLASMINOGEN IN VIBRIO CHOLERAE


Cite item

Full Text

Abstract

Aim. Study system of activation of plasminogen in Vibrio cholerae. Materials and methods. 75 strains of V. cholerae of various origins were used in the study. Plasminogen was isolated from human plasma by using affinity chromatography on L-lysine sepharose, α-enolase activity was determined by a direct method assuming transformation of 2-phosphoglycerate into phopshoenolpyruvate. Vibrios were destroyed by ultrasound disintegrator to isolate membrane OmpT protein, intact cells were discarded by centrifugation and cell lysate was centrifugated for 1 hour at 105000 g. The precipitate was solubilized in buffer with 1% triton X-100 and passed through a column with DE-52 cellulose. Results. Vibrio cholerae O1 and O139 strains isolated from clinical specimens and water samples from open water bodies had the ability to bind by using α-enolase and transform human plasminogen into plasmin under the effect ofouter membrane protein OmpT. A protein with molecular weight around 40 kDa had proteolytic activity with a wide specter of substrate specificity, degraded fibrin, gelatin, collagen, protamine and activated plasminogen. Computer analysis showed that OmpT protein of cholera vibrion had a low degree of relation with Enterobacteriaceae omptins. Conclusion. The study carried out showed that vibrios have a system of activation of plasminogen that includes at least α-enolase and OmpT membrane protein. OmpT protein is assumed to belong to a new class of porins of Vibrionaceae family and its enzymatic activity may play a significant role in pathogenesis of infection.

About the authors

B. N Mishankin

Rostov-on-Don Institute of Plague Control, Russia

O. V Duvanova

Rostov-on-Don Institute of Plague Control, Russia

E. S Shipko

Rostov-on-Don Institute of Plague Control, Russia

L. V Romanova

Rostov-on-Don Institute of Plague Control, Russia

A. S Vodopianov

Rostov-on-Don Institute of Plague Control, Russia

A. K Eribekyan

Rostov-on-Don Institute of Plague Control, Russia

M. V Shishiyanu

Rostov-on-Don Institute of Plague Control, Russia

L. K Lysova

Rostov-on-Don Institute of Plague Control, Russia

S. O Vodopianov

Rostov-on-Don Institute of Plague Control, Russia

References

  1. Ашмарин И.П., Воробьев А.А. Статистические методы в микробиологических исследованиях. Л., 1962.
  2. Миронова А.В., Меньшикова Е.А. Инвазивность токсигенных и нетоксигенных холерных вибрионов различных серогрупп. Журн. микробиол. 2002, 3: 58-60.
  3. Мишанькин Б.Н., Гончаров Е.К., Марченков В.И. и др. Молекулярное клонирование плазмиды Yersinia pestis, детерминирующей синтез пестицина, белка иммунности, фибриноли-зина и коагулазы. Молекулярная генетика, микробиология и вирусология. 1984, 10: 12-16.
  4. Рыжко И.В., Цураева Р.И., Ломов Ю.М. и др. Результаты оценки активности антибактериальных препаратов in vitro и in vivo эпидемически значимых штаммов холерного вибриона биотипа эльтор. Эпидемиология и инфекционные болезни. 2000, 3: 21-25.
  5. Шипко Е.С., Мишанькин Б.Н., Дуванова О.В. и др. Плазминоген-активирующая способность холерного вибриона и участие мембранного белка OmpT в этом процессе. Защита населения и среда обитания. 2012, 4: 17-19.
  6. Bergmann S., Wild D., Dickmann O. et al. Identification of a novel plasmin(ogen) - binding motif in surface displayed α-enolase of Streptococcus pneumoniae. Mol. Microbiol. 2003, 40: 411-423.
  7. Candela M., Miccoli G., Bergmann S. et al. Plasminogen - dependent proteolytic activity in Bifidobacterium lactis. Microbiology. 2008, 153: 2457-2462.
  8. Chakrabarti S.R., Chaudhuri K., Sen K. et al. Porins of Vibrio cholerae: purification and characterization of OmpU. J. Bacteriol. 1996, 178 (2): 534-538.
  9. Deutsch D.G., Mertz E.T. Plasminogen: purification from human plasma by affinity chromatography Science. 1970, 170: 1095-1096.
  10. Gordon M.A., Walsh A.L., Rogerson S.R.K. et al. Three cases of bacteremia caused by Vibrio cholerae O1 in Blantyre, Malawi. Emerg. Infect. Diseases. 2001, 7 (6): 1059-1061.
  11. Haiko J., Suomalainen M., Ojala T. et al. Invited review: Breaking barrier-attack on innate immune defences by omptin surface proteases of enterobacterial pathogens. Innate Immun. 2009, 15 (2): 67-80.
  12. Lahteenmaki K., Edelman S., Korhonen T.K. Bacterial metastasis: the host plasminogen system in bacterial invasion. Trends in Microbiology. 2005, 13 (2): 79-85.
  13. Merrell D.S., Bailey C., Kaper J.B. et al. The ToxR-mediated organic acid tolerance response of Vibrio cholerae requires OmpU. J. Bacteriol. 2001, 183 (9): 2746-2754.
  14. Neutra M.R., Frey A., Kraehenbuhl J.P. Epithelial M-cells: gateways for mucosal infection and immunization. Cell. 1996, 86: 345-348.
  15. Pancholi V, Fischetti V. Alpha-enolase, a novel strong plasmin(ogen) binding protein on the surface of pathogenic streptococci. J. Biol. Chem. 1998, 273 (23): 14503-14515.
  16. Pouillot F., Derbise A., Kukkonen M. et al. Evaluation of O-antigen inactivation on Pla activity and virulence ofYersinia pseudotuberculosis harbouring the pPla plasmid. Microbiology. 2005, 151: 3759-3768.
  17. Sha J., Erova T.E., Alyea R.A. et al. Surface-expressed enolase contributes to the pathogenesis of clinical isolate SSU of Aeromonas hydrophila. J. Bacteriol. 2009, 191 (9): 3095-3107.
  18. Stumpe S., Schmid R., Stefens D.L. et al. Identification of OmpT as the protease that hydrolyzes the antimicrobial peptide protamine before it enters growing cells of E. coli. J. Bacteriol. 1998, 180: 4002-4006.
  19. Zhang L., Seiffert D., Fowler B.J. et al. Plasminogen has a broad extrahepatic distribution. Tromb. Haemost. 2002, 87 (3): 493-501.

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2013 Mishankin B.N., Duvanova O.V., Shipko E.S., Romanova L.V., Vodopianov A.S., Eribekyan A.K., Shishiyanu M.V., Lysova L.K., Vodopianov S.O.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: ПИ № ФС77-75442 от 01.04.2019 г.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies