Fibrinogen and Fibronectin Binding Activity and Immunogenic Nature of Choline Binding Protein M
-
Davoud AFSHAR
,
Mohammad Reza POURMAND
,
Mahmood JEDDI-TEHRANI
,
Ali Akbar SABOOR YARAGHI
,
Mohammad AZARSA
,
Fazel SHOKRI
Abstract
Background: Choline-binding proteins (CBPs) are a group of surface-exposed proteins, which play crucial and physiological roles in Streptococcus pneumoniae. The novel member of CBPs, choline-binding protein M (CbpM) may have binding activity to plasma proteins. This study aimed to clone and express CbpM and demonstrate its interaction with plasma proteins and patients’ sera.
Methods: The total length of cbpM gene was cloned in pET21a vector and expressed in BL21 expression host. Verification of recombinant protein was evaluated by Western blot using anti-His tag monoclonal antibody. Binding ability of the recombinant protein to plasma proteins and the interaction with patients’ sera were assessed by Western blot and ELISA methods.
Results: The cbpM gene was successfully cloned into pET21a and expressed in BL21 host. Binding activity to fibronectin and fibrinogen and antibody reaction of CbpM to patients’ sera was demonstrated by Western blot and ELISA methods, respectively.
Conclusion: CbpM is one of the pneumococcal surface-exposed proteins, which mediates pneumococcal binding to fibronectin and fibrinogen proteins.
Jedrzejas MJ (2001). Pneumococcal virulence factors: structure and function. Microbiol Mol Biol Rev, 65(2):187-207.
Yother J, Leopold K, White J et al (1998). Generation and properties of a Streptococcus pneumoniae mutant, which does not require choline or analogs for growth. J Bacteriol, 180(8):2093-101.
Moscoso M, Garcia E, Lopez R (2006). Biofilm formation by Streptococcus pneumoniae: role of choline, extracellular DNA, and capsular polysaccharide in microbial accretion. J Bacteriol, 188(22):7785-95.
Kausmally L, Johnsborg O, Lunde M et al (2005). Choline-binding protein D (CbpD) in Streptococcus pneumoniae is essential for competence-induced cell lysis. J Bacteriol, 187(13):4338-45.
Feldman C, Anderson R (2014). Review: current and new generation pneumococcal vaccines. J Infect, 69(4):309-25.
Luo R, Mann B, Lewis WS et al (2005). Solution structure of choline binding protein A, the major adhesin of Streptococcus pneumoniae. EMBO J, 24(1):34-43.
Orihuela CJ, Gao G, Francis KP et al (2004). Tissue-specific contributions of pneumococcal virulence factors to pathogenesis. J Infect Dis, 190(9):1661-69.
Ogunniyi AD, Woodrow MC, Poolman JT et al (2001). Protection against Streptococcus pneumoniae elicited by immunization with pneumolysin and CbpA. Infect Immun, 69(10):5997-6003.
Iannelli F, Oggioni MR, Pozzi G (2002). Allelic variation in the highly polymorphic locus pspC of Streptococcus pneumoniae. Gene, 284(1):63-71.
Gosink KK, Mann ER, Guglielmo C et al (2000). Role of novel choline binding proteins in virulence of Streptococcus pneumoniae. Infect Immun, 68 (10):5690-95.
Hammerschmidt S (2006). Adherence molecules of pathogenic pneumococci. Curr Opin Microbiol, 9(1):12-20.
Mann B, Orihuela C, Antikainen J et al (2006). Multifunctional role of choline binding protein G in pneumococcal pathogenesis. Infect Immun, 74 (2):821-29.
Frolet C, Beniazza M, Roux L et al (2010). New adhesin functions of surface-exposed pneumococcal proteins. BMC Microbiol, 10(1):190.
Ramina Mahboobi, Davoud Afshar, Mohammad Reza Pourmand, Rahil Mashhadi (2016). Autolytic activity and plasma binding study of Aap, a novel minor autolysin of Streptococcus pneumoniae. Acta Med Iran, 54(3):196-200.
Marchler-Bauer A, Derbyshire MK, Gonzales NR et al (2015). CDD: NCBI's conserved domain database. Nucleic Acids Res, 43:D222-26.
Bateman A, Rawlings ND (2003). The CHAP domain: a large family of amidases including GSP amidase and peptidoglycan hydrolases. Trends Biochem Sci, 28(5):234-37.
Mitchell J, Tristan A, Foster TJ (2004). Characterization of the fibrinogen-binding surface protein Fbl of Staphylococcus lugdunensis. Microbiology, 150(11):3831-41.
Shannon O, Flock JI (2004). Extracellular fibrinogen binding protein, Efb, from Staphylococcus aureus binds to platelets and inhibits platelet aggregation. Thromb Haemost, 91(4):779-89.
Hasty D, Ofek I, Courtney H, Doyle R (1992). Multiple adhesins of streptococci. Infect Immun, 60(6):2147-52.
Molinari G, Rohde M, Guzmán CA, Chhatwal GS (2000). Two distinct pathways for the invasion of Streptococcus pyogenes in non‐phagocytic cells. Cell Microbiol, 2(2):145-54.
Bumbaca D, LittleJohn JE, Nayakanti H, Rigden DJ, Galperin MY, Jedrzejas MJ (2004). Sequence analysis and characterization of a novel fibronectin-binding repeat domain from the surface of Streptococcus pneumoniae. Omics, 8(4):341-56.
Holmes AR, McNab R, Millsap KW, Rohde M, Hammerschmidt S, Mawdsley JL, et al (2001). The pavA gene of Streptococcus pneumoniae encodes a fibronectin‐binding protein that is essential for virulence. Mol Microbiol, 41(6):1395-408.
Voss S, Hallstrom T, Saleh M et al (2013). The choline-binding protein PspC of Streptococcus pneumoniae interacts with the C-terminal heparin-binding domain of vitronectin. J Biol Chem, 288(22):15614-27.
Bergmann S, Lang A, Rohde M et al (2009). Integrin-linked kinase is required for vitronectin-mediated internalization of Streptococcus pneumoniae by host cells. J Cell Sci, 122(2):256-67.
Kelley LA, Sternberg MJ (2009). Sternberg MJ Protein structure prediction on the Web: a case study using the Phyre server. Nat Protoc, 4(3):363-71.
| Files | ||
| Issue | Vol 45 No 12 (2016) | |
| Section | Original Article(s) | |
| Keywords | ||
| Streptococcus pneumoniae Choline-binding protein M Fibrinogen Fibronectin | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
