Ferritin Degradation by Pneumococcal HtrA, RadA and ClpP Serine Proteases : A Probable Way For Releasing and Acquisition Of Iron.

Iron is a necessary element for the growth of bacteria; however, there are limited iron sources known for these microorganisms yet. Intracellular iron is stored as ferritin from, which releases iron in a gradual and controlled manner. The present study aimed to characterize ferritin-binding proteins (FBPs) of Streptococcus pneumoniae. S. pneumoniae species were cultured in BHI broth containing ferritin (1094 ng/mL) for 4h at 37°C. Ferritin level was measured using ELISA assay. Bacterial proteome was electrophoresed on SDS-PAGE and then transferred on PVDF nitrocellulose membrane. Afterward, the PVDF membranes were incubated with a ferritin solution. Identification of ferritin binding proteins was performed using anti-ferritin monoclonal antibody conjugated with HRP enzyme. Molecular docking was used to assess the interaction between pneumococcal proteases and FBPs applying phenylmethylsulfonyl fluoride (PMSF) as a protease inhibitor. No FBPs were identified in S. pneumoniae proteome. Moreover, ferritin levels have significantly (p<0.05) decreased following the growth of S. pneumoniae in ferritin-rich BHI medium. Also, molecular docking showed that RadA protease, ClpP hydrolase, and HtrA protease can potentially interact with PMSF protease inhibitors. On the other hand, the addition of the PMSF to the culture of S. pneumoniae prevented the reduction of ferritin, which indicates a potential role of RadA, ClpP, and HtrA proteases in ferritin degradation. Our results suggest that S. pneumoniae produces no FBPs and also cannot directly use ferritin as an iron source. However, ferritin may be degraded through a protease-mediated mode. © 2020 Kheirandish et al.

Citation

Mozhgan Kheirandish, Behrooz Motlagh, Davoud Afshar. Ferritin Degradation by Pneumococcal HtrA, RadA and ClpP Serine Proteases : A Probable Way For Releasing and Acquisition Of Iron. Infection and drug resistance. 2020;13:3145-3152

PMID: 32982329

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