Abstract
Streptococcus pyogenes harboring an FCT type 3 genomic region display pili composed of three types of pilins. In this study, the structure of the base pilin FctB from a serotype M3 strain (FctB3) was determined at 2.8 Å resolution. In accordance with the previously reported structure of FctB from a serotype T9 strain (FctB9), FctB3 was found to consist of an immunoglobulin-like domain and proline-rich tail region. Data obtained from structure comparison revealed main differences in the omega (Ω) loop structure and the proline-rich tail direction. In the Ω loop structure, a differential hydrogen bond network was observed, while the lysine residue responsible for linkage to growing pili was located at the same position in both structures, which indicated that switching of the hydrogen bond network in the Ω loop without changing the lysine position is advantageous for linkage to the backbone pilin FctA. The difference in direction of the proline-rich tail is potentially caused by a single residue located at the root of the proline-rich tail. Also, the FctB3 structure was found to be stabilized by intramolecular large hydrophobic interactions instead of an isopeptide bond. Comparisons of the FctB3 and FctA structures indicated that the FctA structure is more favorable for linkage to FctA. In addition, the heterodimer formation of FctB with Cpa or FctA was shown to be mediated by the putative chaperone SipA. Together, these findings provide an alternative FctB structure as well as insight into the interactions between pilin proteins.
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Data availability
The protein structure data are available in the wwPDB repository under accession number 8K6T.
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Acknowledgements
We thank T. Sekizaki (Kyoto University) and D. Takamatsu (National Institute of Animal Health) for providing the pSET4s plasmid. pAT18 was kindly provided by P. Trieu-Cuot (Institut Pasteur). This work was performed using a synchrotron beamline BL44XU at SPring-8 under the Collaborative Research Program of Institute for Protein Research, Osaka University. Preliminary diffraction data were collected at the Osaka University beamline BL44XU at SPring-8 (Harima, Japan) (Proposal No. 2017A6732, 2017B6732, and 2018A6832). Also, this work was performed under the approval of the Photon Factory Program Advisory Committee (Proposal No. 2016G609, 2017G169).
Funding
This work was supported by JSPS KAKENHI Grants‐in‐Aid for Scientific Research (Grant Nos. 19K22715, 19H03825, 22H03262, 22H03263).
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KT and MN contributed to the study conception and design. Material preparation, data collection, and analysis were performed by KT, MS, TS, and MN. The first draft of the manuscript was written by KT and MN, and all authors contributed to revision of the manuscript. All authors approved the final manuscript.
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Takebe, K., Suzuki, M., Sangawa, T. et al. Analysis of FctB3 crystal structure and insight into its structural stabilization and pilin linkage mechanisms. Arch Microbiol 206, 4 (2024). https://doi.org/10.1007/s00203-023-03727-1
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DOI: https://doi.org/10.1007/s00203-023-03727-1