Fibrinogen and Fibronectin Binding Activity and Immunogenic Nature of Choline Binding Protein M
Davoud Afshar1, Mohammad Reza Pourmand2, Mahmood Jeddi-Tehrani3
1Dept. of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Insights
Choline-binding protein M (CbpM) from Streptococcus pneumoniae binds to human fibronectin and fibrinogen. This study successfully cloned and expressed CbpM, confirming its role in bacterial adhesion.
Area of Science:
- Microbiology
- Molecular Biology
- Protein Biochemistry
Background:
- Choline-binding proteins (CBPs) are essential surface proteins in *Streptococcus pneumoniae*, involved in critical physiological functions.
- CbpM, a novel CBP, is investigated for its potential to bind plasma proteins, suggesting a role in host-pathogen interactions.
Purpose of the Study:
- To clone and express the choline-binding protein M (CbpM) from *Streptococcus pneumoniae*.
- To investigate the binding activity of CbpM to plasma proteins and its interaction with human patient sera.
Main Methods:
- The *cbpM* gene was cloned into the pET21a vector and expressed in the BL21 host.
- Recombinant CbpM protein was verified using Western blot with an anti-His tag antibody.
- Binding to plasma proteins and interaction with patient sera were assessed via Western blot and ELISA.
Main Results:
- Successful cloning and expression of the *cbpM* gene and recombinant CbpM protein.
- Demonstrated binding activity of CbpM to fibronectin and fibrinogen.
- Confirmed antibody reaction of CbpM to patient sera, indicating potential immunogenic properties.
Conclusions:
- CbpM is identified as a pneumococcal surface protein involved in binding to host fibronectin and fibrinogen.
- These findings elucidate a mechanism for *Streptococcus pneumoniae* adhesion to host tissues via CbpM.
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.
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