Abstract
Osteoblasts produce the receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin, the inducer and the suppressor of osteoclast differentiation and activation. We previously proposed that the degradation of osteoprotegerin by lysine-specific gingipain of Porphyromonas gingivalis and neutrophil elastase is one of the mechanisms of bone resorption associated with infection and inflammation. In the present study, we found that cathepsin K (CTSK) also degraded osteoprotegerin in an acidic milieu and the buffer with a pH of 7.4. The 37 k fragment of osteoprotegerin produced by the reaction with CTSK was further degraded into low molecular weight fragments, including a 13 k fragment, depending on the reaction time. The N-terminal amino acid sequence of the 37 k fragment matched that of the intact osteoprotegerin, indicating that CTSK preferentially hydrolyzes the death domain-like region of osteoprotegerin, not its RANKL-binding region. The 13 k fragment of osteoprotegerin was the C-terminal 13 k portion within the RANKL-binding region of the 37 k fragment. Finally, CTSK restored RANKL-dependent osteoclast differentiation that was suppressed by the addition of osteoprotegerin. Collectively, CTSK is a possible positive regulator of osteoclastogenesis.
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The datasets of the current study are available from the corresponding author upon reasonable request.
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Funding
This work was supported by Grants-In-Aid for Scientific Research (KAKENHI) from the Japan Society for the Promotion of Science (21K09835, 22H03256).
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Kawai, Sugisaki, Yano, Sasa, and Minami performed experiments. Miyamoto designed the study. Maki and Kamijo analyzed the data. Kawai and Miyamoto prepared the original draft of the manuscript. Miyamoto, Maki, and Kamijo reviewed and edited the manuscript. All authors have read and agreed with the final version of the manuscript.
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Kawai, R., Sugisaki, R., Miyamoto, Y. et al. Cathepsin K degrades osteoprotegerin to promote osteoclastogenesis in vitro. In Vitro Cell.Dev.Biol.-Animal 59, 10–18 (2023). https://doi.org/10.1007/s11626-023-00747-5
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DOI: https://doi.org/10.1007/s11626-023-00747-5