Negative regulation of osteoclastogenesis by ectodomain shedding of receptor activator of NF-κB ligand
Negative regulation of osteoclastogenesis by ectodomain shedding of receptor activator of NF-κB ligand
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DOI：
10.1074/jbc.m606656200
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发表时间：
2006-12-01
影响因子：
4.8
通讯作者：
Tanaka, Sakae
中科院分区：
文献类型：
--
作者：
Hikita, Atsuhiko;Yana, Ikuo;Tanaka, Sakae
nf - κ B配体受体激活因子（Receptor activator of nf - κ B ligand， RANKL）是一种跨膜糖蛋白，在破骨细胞的发育中起重要作用。RANKL的细胞外部分被蛋白水解裂解以产生可溶性RANKL，但明确的RANKL脱落酶和RANKL脱落的生理功能尚未确定。在本研究中，我们发现基质金属蛋白酶（MMP） 14和崩解素和金属蛋白酶（ADAM） 10具有较强的RANKL脱落活性。在Western blot分析中，可溶性RANKL被检测为两种不同分子量的产物，MMP14和ADAM10的RNA干扰导致低分子量和高分子量产物的减少。在与巨噬细胞共培养中，抑制原代成骨细胞中的MMP14增加了膜结合RANKL并促进了破骨细胞的形成。mmp14缺失小鼠的成骨细胞产生的可溶性RANKL显著减少，其破骨活性增强，与体内破骨细胞生成增加的结果一致。RANKL脱落是下调局部破骨细胞发生的重要过程。
Receptor activator of NF-kappa B ligand (RANKL) is a transmembrane glycoprotein that has an essential role in the development of osteoclasts. The extracellular portion of RANKL is cleaved proteolytically to produce soluble RANKL, but definite RANKL sheddase(s) and the physiologic function of RANKL shedding have not yet been determined. In the present study, we found that matrix metalloproteinase (MMP) 14 and a disintegrin and metalloproteinase (ADAM) 10 have strong RANKL shedding activity. In Western blot analysis, soluble RANKL was detected as two different molecular weight products, and RNA interference of MMP14 and ADAM10 resulted in a reduction of both the lower and higher molecular weight products. Suppression of MMP14 in primary osteoblasts increased membrane-bound RANKL and promoted osteoclastogenesis in cocultures with macrophages. Soluble RANKL produced by osteoblasts from MMP14-deficient mice was markedly reduced, and their osteoclastogenic activity was promoted, consistent with the findings of increased osteoclastogenesis in vivo. RANKL shedding is an important process that down-regulates local osteoclastogenesis.