Department of Biochemistry, Showa University School of Dentistry, Tokyo, Japan.
Department of Orthodontics, Showa University School of Dentistry, Tokyo, Japan.
In Vitro Cell Dev Biol Anim. 2023 Jan;59(1):10-18. doi: 10.1007/s11626-023-00747-5. Epub 2023 Jan 23.
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.
成骨细胞产生核因子-κ B 配体(RANKL)的受体激活剂和骨保护素,骨保护素是破骨细胞分化和激活的诱导剂和抑制剂。我们之前提出,牙龈卟啉单胞菌的赖氨酸特异性牙龈蛋白酶和中性粒细胞弹性蛋白酶对骨保护素的降解是与感染和炎症相关的骨吸收的机制之一。在本研究中,我们发现组织蛋白酶 K(CTSK)也可以在酸性环境中和 pH 值为 7.4 的缓冲液中降解骨保护素。与 CTSK 反应产生的骨保护素 37k 片段进一步降解为低分子量片段,包括 13k 片段,这取决于反应时间。37k 片段的 N 末端氨基酸序列与完整的骨保护素匹配,表明 CTSK 优先水解骨保护素的死亡结构域样区域,而不是其 RANKL 结合区域。骨保护素的 13k 片段是 37k 片段的 RANKL 结合区域内的 C 末端 13k 部分。最后,CTSK 恢复了 RANKL 依赖性破骨细胞分化,这种分化被添加骨保护素所抑制。总之,CTSK 可能是破骨细胞生成的正调节剂。