Fibrinogen and mucin binding activity of EF0737, a novel protein of Enterococcus faecalis

  • Samira Sabzi Department of Pathobiology, School of Public Health and Biotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Rahil Mashhadi Urology Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Reza Pourmand Department of Pathobiology, School of Public Health and Biotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran
Keywords: Enterococcus faecalis, EF0737, Fibrinogen, Mucin, Pathogenesis


Background and Objectives: Enterococcus faecalis is the leading cause of several human infections. This opportunist pathogen expresses surface components that have various functions in the infection process including bacterial adhesion, lytic activity, and induction of host immune responses. EF0737, a novel cell wall associated protein, may play an important role in pathogenesis of E. faecalis, based on our experiments. This study was conducted to clone and express EF0737 and demonstrate its interaction with biotinylated plasma proteins and patients’ sera.Materials and Methods: The full length of ef0737 gene was cloned in pTZ57R/T cloning vector and subcloned in pET21a expression vector. Recombinant protein expressed in Escherichia coli Origami (DE3) was confirmed by western blot technique, using anti-His tagged monoclonal antibodies, and was then purified. Interaction of the recombinant protein with plasma proteins and patients’ sera were examined by western blot.Results: The ef0737 gene was successfully cloned and expressed in E. coli Origami host. Binding activity was observed between the purified EF0737 recombinant protein and fibrinogen and mucin among other plasma proteins. Moreover, reaction was also observed between the purified product and sera obtained from patients diagnosed with E. faecalis infection.Conclusion: The observed reactions between EF0737 and fibrinogen, mucin and patients’ sera suggest that EF0737 may play important role in pathogenesis of infections caused by E. faecalis. However, more comprehensive characterization of this novel protein may provide better understanding of host pathogen interaction.


Guiton PS, Hung CS, Kline KA, Roth R, Kau AL, Hayes E, et al. Contribution of autolysin and Sortase a during Enterococcus faecalis DNA-dependent biofilm development. Infect Immun 2009;77:3626-3638.

AgudeloHiguita NI, Huycke MM (2014). Enterococcal disease, epidemiology, and implications for treatment. In: From Commensals to Leading Causes of Drug Resistant Infection.Eds M. S. Gilmore, D. B. Clewell, Y. Ike, N. Shankar. Massachusetts Eye and Ear Infirmary, Boston, pp.1-27.

Hollenbeck BL, Rice LB. Intrinsic and acquired resistance mechanisms in enterococcus. Virulence 2012;3:421-433.

Toledo-Arana A, Valle J, Solano C, ArrizubietaMa J, Cucarella C, Lamata M, et al. The enterococcal surface protein, Esp, is involved in Enterococcus faecalis biofilm formation. Appl Environ Microbiol 2001;67(10):4538-4545.

Somarajan SR, La Rosa SL, Singh KV, Roh JH, Höök M, Murray BE. The fibronectin binding protein fnm contributes to adherence to extracellular matrix components and virulence of Enterococcus faecium. Infect Immun 2015;83:4653-4561.

Romero-Saavedra F, Laverde D, Wobser D, Michaux C, Budin-Verneuil A, Bernay B, et al. Identification of peptidoglycan-associated proteins as vaccine candidates for enterococcal infections. PLoS One 2014;9(11):e111880.

Singh KV, La Rosa SL, Somarajan SR, Roh JH, Murray BE. The fibronectin-binding protein EfbA contributes to pathogenesis and protects against infective endocarditis caused by Enterococcus faecalis. Infect Immun 2015;83:4487-4494.

Sillanpää J, Nallapareddy SR, Houston J, Ganesh VK, Bourgogne A, Singh KV, et al. A family of fibrinogen-binding MSCRAMMs from Enterococcus faecalis. Microbiology 2009;155(Pt 7):2390-2400.

Bøhle LA, Riaz T, Egge-Jacobsen W, Skaugen M, Busk ØL, Eijsink VG, et al. Identification of surface proteins in Enterococcus faecalis V583. BMC Genomics 2011;12:135.

Heilmann C, Hartleib J, Hussain MS, Peters G. The multifunctional Staphylococcus aureus autolysin aaa mediates adherence to immobilized fibrinogen and fibronectin. Infect Immun 2005;73:4793-4802.

Mellroth P, Daniels R, Eberhardt A, Rönnlund D, Blom H, Widengren J, et al. LytA, major autolysin of Streptococcus pneumoniae, requires access to nascent peptidoglycan. J Biol Chem 2012;287:11018-11029.

Pourmand MR, Clarke SR, Schuman RF, Mond JJ, Foster SJ. Identification of antigenic components of Staphylococcus epidermidis expressed during human infection. Infect Immun 2006;74:4644-4654.

Muscholl-Silberhorn A. Cloning and functional analysis of Asa373, a novel adhesin unrelated to the other sex pheromone plasmid-encoded aggregation substances of Enterococcus faecalis. Mol Microbiol 1999;34:620-630.

Gilmore MS, Coburn PS, Nallapareddy SR, Murray BE (2002). Enterococcal Virulence. In: The Enterococci: Pathogenesis, Molecular Biology, and Antibiotic Resistance. Ed, Gilmore MS, ASM publishing, Washington, 1st ed, pp. 301-54.

Sahay A, Shakya M. In silico analysis and homology modelling of antioxidant proteins of spinach. J Proteomics Bioinform 2010;3:148-54.

Hymes JP, Klaenhammer TR. Stuck in the Middle: Fibronectin-Binding Proteins in Gram-Positive Bacteria. Front Microbiol 2016;7:1504.

Pian Y, Li X, Zheng Y, Wu X, Yuan Y, Jiang Y. Binding of human fibrinogen to MRP enhances Streptococcus suis survival in host blood in a αXβ2 integrin-dependent manner. Sci Rep 2016;6:26966.

Afshar D, PourmandMr, Jeddi-Tehrani M, YaraghiAas, Azarsa M, Shokri F. Fibrinogen and fibronectin binding activity and immunogenic nature of choline binding protein M. Iran J Public Health 2016;45:1610-1617.

Rivera J, Vannakambadi G, Hook M, Speziale P. Fibrinogen-binding proteins of Gram-positive bacteria. Thromb Haemost 2007;98(3):503-511.

Flores-Mireles AL, Pinkner JS, Caparon MG, Hultgren SJ. EbpA vaccine antibodies block binding of Enterococcus faecalis to fibrinogen to prevent catheter-associated bladder infection in mice. Sci Transl Med 2014;6(254):254ra127.

Cornick S, Tawiah A, Chadee K. Roles and regulation of the mucus barrier in the gut. Tissue Barriers 2015;3(1-2):e982426.

Ryan PA, Pancholi V, Fischetti VA. Group A streptococci bind to mucin and human pharyngeal cells through sialic acid-containing receptors. Infect Immun 2001;69(12):7402-7412.

How to Cite
Sabzi S, Mashhadi R, Pourmand M. Fibrinogen and mucin binding activity of EF0737, a novel protein of Enterococcus faecalis. IJM. 9(6):324-30.
Original Article(s)