Negative regulation of osteoclastogenesis by ectodomain shedding of receptor activator of NF-kappaB ligand.
Receptor activator of NF-kappaB 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.
Hikita
Atsuhiko
A
Department of Orthopaedic Surgery, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, USA.
Yana
Ikuo
I
Wakeyama
Hidetoshi
H
Nakamura
Masaki
M
Kadono
Yuho
Y
Oshima
Yasushi
Y
Nakamura
Kozo
K
Seiki
Motoharu
M
Tanaka
Sakae
S
D051722:ADAM Proteins / Q000737:chemistry; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D001851:Bone Diseases, Metabolic / Q000378:metabolism; D001853:Bone Marrow / Q000378:metabolism; D018920:Coculture Techniques; D015536:Down-Regulation; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D010006:Osteoblasts / Q000378:metabolism; D010010:Osteoclasts / Q000378:metabolism*; D053245:RANK Ligand / Q000737:chemistry*
D008565:Membrane Proteins; D053245:RANK Ligand; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D053511:Matrix Metalloproteinase 14; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; C491234:Adam10 protein, mouse