Characterization of Afb, a novel bifunctional protein in Streptococcus agalactiae
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Abstract
Background and Objectives: Streptococcus agalactiae is the leading cause of bacterial sepsis and meningitis in newborns and results in pneumonia and bacteremia in adults. A number of S. agalactiae components are involved in colonization of target cells. Destruction of peptidoglycan and division of covalently linked daughter cells is mediated by autolysins. In this study, autolytic activity and plasma binding ability of AFb novel recombinant protein of S. agalactiae was investigated.
Materials and Methods: The gbs1805 gene was cloned and expressed. E. coli strains DH5α and BL21 were used as cloning and expression hosts, respectively. After purification, antigenicity and binding ability to plasma proteins of the recombinant protein was evaluated.
Results: AFb, the 18KDa protein was purified successfully. The insoluble mature protein revealed the ability to bind to fibrinogen and fibronectin. This insoluble mature protein revealed that it has the ability to bind to fibrinogen and fibronectin plasma proteins. Furthermore, in silico analysis demonstrated the AFb has an autolytic activity.
Conclusions: AFb is a novel protein capable of binding to fibrinogen and fibronectin. This findings lay a ground work for further investigation of the role of the bacteria in adhesion and colonization to the host.
Reinscheid DJ, Gottschalk B, Schubert A, Eikmanns BJ, Chhatwal GS. Identification and molecular analysis of PcsB, a protein required for cell wall separation of group B streptococcus. J Bacteriol 2001; 183:1175-83.
Ragunathan P, Ponnuraj K. Expression, purification and structural analysis of a fibrinogen receptor FbsA from Streptococcus agalactiae. Protein J 2011; 30;159-66.
Fluegge K, Schweier O, Schiltz E, Batsford S, Berner R. Identification and immunoreactivity of proteins re- leased from Streptococcus agalactiae. Eur J Clin Mi- crobiol Infect Dis 2004; 23: 818-24.
Hull JR, Tamura GS, Castner DG. Interactions of the streptococcal C5a peptidase with human fibronectin. Acta Biomater 2008; 4: 504-513.
Butler KM, Baker CJ, Edwards MS. Interaction of sol- uble fibronectin with group B streptococci. Infect Im- mun 1987; 55: 2404-2408.
Beckmann C, Waggoner JD, Harris TO, Tamura GS, Rubens CE. Identification of novel adhesins from Group B streptococci by use of phage display reveals that C5a peptidase mediates fibronectin binding. Infect Immun 2002; 70: 2869-2876.
Rivera J, Vannakambadi G, Hook M, Speziale P. Fi- brinogen-binding proteins of Gram-positive bacteria. Thromb Haemost 2007; 98: 503-511.
Devi AS, Ponnuraj K. Cloning, expression, purification and ligand binding studies of novel fibrinogen-binding protein FbsB of Streptococcus agalactiae. Protein Expr Purif 2010; 74: 148-155.
Gutekunst H, Eikmanns BJ, Reinscheid DJ. The novel fibrinogen-binding protein FbsB promotes Streptococ- cus agalactiae invasion into epithelial cells. Infect Im- mun 2004; 72: 3495-504.
Gotz F, Heilmann C, Stehle T. Functional and structur- al analysis of the major amidase (Atl) in Staphylococ- cus. Int J Med Microbiol 2014; 304: 156-63.
Biswas R, Voggu L, Simon UK, Hentschel P, Thumm G, Gotz F. Activity of the major staphylococcal autoly- sin Atl. FEMS Microbiol Lett 2006; 259: 260-268.
Garcia P, Paz GM, Garcia E, Garcia JL, Lopez R. The molecular characterization of the first autolytic lyso- zyme of Streptococcus pneumoniae reveals evolution- ary mobile domains. Mol Microbiol 1999; 33:128-138.
Grilo IR, Ludovice AM, Tomasz A, De LencastreH, So- bral RG. The glucosaminidase domain of Atl - the ma- jor Staphylococcus aureus autolysin - has DNA-bind- ing activity. Microbiology open 2014; 3: 247-256.
Heilmann C, Hartleib J, Hussain MS, Peters G. The multifunctional Staphylococcus aureus autolysin aaa mediates adherence to immobilized fibrinogen and fi- bronectin. Infect Immun 2005; 73: 4793-4802.
Mellroth P, Daniels R, Eberhardt A, Ronnlund D, Blom H, Widengren J, et al. LytA, major autolysin of Strep- tococcus pneumoniae, requires access to nascent pepti- doglycan. J Biol Chem 2012; 287: 11018-29.
Tamura GS, Kuypers JM, Smith S, Raff H, Rubens CE.Adherence of group B streptococci to cultured epithe- lial cells: roles of environmental factors and bacterial surface components. Infect Immun 1994; 62: 2450-2458.
Patti JM, Allen BL, McGavin MJ, Hook M.MSCRAMM-mediated adherence of microorganisms to host tissues. Annu Rev Microbiol 1994; 48:585-617.
Courtney HS, Bronze MS, Dale JB, Hasty DL. Analy- sis of the role of M24 protein in group A streptococcal adhesion and colonization by use of omega-interposon mutagenesis. Infect Immun 1994; 62: 4868-4873.
Shenkman B, Rubinstein E, Tamarin I, Dardik R, Sav- ion N, Varon D. Staphylococcus aureus adherence to thrombin-treated endothelial cells is mediated by fi- brinogen but not by platelets. J Lab Clin Med 2000;135: 43-51.
Buscetta M, Papasergi S, Firon A, Pietrocola G, Biondo C, Mancuso G, et al. FbsC, a Novel Fibrinogen-binding Protein, Promotes Streptococcus agalactiae-Host Cell Interactions. J Biol Chem 2014; 289: 21003-21015.
Seo HS, Minasov G, Seepersaud R, Doran KS, Du- brovska I, Shuvalova L, et al. Characterization of fi- brinogen binding by glycoproteins Srr1 and Srr2 of Streptococcus agalactiae. J Biol Chem 2013; 288:35982-35996.
Schubert A, Zakikhany K, Schreiner M, Frank R,Spellerberg B, Eikmanns BJ, et al. A fibrinogen recep-tor from group B Streptococcus interacts with fibrino- gen by repetitive units with novel ligand binding sites. Mol Microbiol 2002; 46: 557-569.
Margarit I, Bonacci S, Pietrocola G, Rindi S, Ghezzo C, Bombaci M, et al. Capturing host-pathogen inter- actions by protein microarrays: identification of novel streptococcal proteins binding to human fibronectin, fibrinogen, and C4BP. FASEB J 2009; 23: 3100-3112.
Seo HS, Xiong YQ, Sullam PM. Role of the serine-rich surface glycoprotein Srr1 of Streptococcus agalactiae in the pathogenesis of infective endocarditis. PLoS One 2013; 8: e64204.
Mu R, Kim BJ, Paco C, Del Rosario Y, Courtney HS, Doran KS. Identification of a group B streptococcal fibronectin binding protein, Sf bA, that contributes to invasion of brain endothelium and development of meningitis. Infect Immun 2014;82: 2276-86.
Berry AM, Paton JC. Additive attenuation of virulence of Streptococcus pneumoniae by mutation of the gene encoding pneumolysin and other putative pneumococ- cal virulence proteins. Infect Immun 2000; 68: 133-40.
Hell W, Meyer HG, Gatermann SG. Cloning of aas, a gene encoding a Staphylococcus saprophyticus surface protein with adhesive and autolytic properties. Mol Mi- crobiol 1998; 29: 871-881.
Milohanic E, Jonquieres R, Cossart P, Berche P, Gail- lard JL. The autolysin Ami contributes to the adhesion of Listeria monocytogenes to eukaryotic cells via its cell wall anchor. Mol Microbiol 2001; 39: 1212-1224.
Heilmann C, Thumm G, Chhatwal GS, Hartleib J, Ue- kotter A, Peters G. Identification and characterization of a novel autolysin (Aae) with adhesive properties from Staphylococcus epidermidis. Microbiology 2003;149: 2769-78.
Rupp ME, Fey PD, Heilmann C, Gotz F. Characteriza- tion of the importance of Staphylococcus epidermidis autolysin and polysaccharide intercellular adhesin in the pathogenesis of intravascular catheter-associated infection in a rat model. J Infect Dis 2001; 183:1038-1042.
Pilgrim S, Kolb-Maurer A, Gentschev I, Goebel W, Kuhn M. Deletion of the gene encoding p60 in Listeria monocytogenes leads to abnormal cell division and loss of actin-based motility. Infect Immun 2003; 71: 3473-84.
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| Issue | Vol 8 No 1 (2016) | |
| Section | Articles | |
| Keywords | ||
| Streptococcus agalactiae Fibrinogen Fibronectin Autolysin | ||
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