Abstract
Osteoclastogenesis is a highly sophisticated process that involves a variety of membrane-bound proteins expressed in osteoblasts and osteoclast precursors. Over the past several years, proteolytic cleavage and release of the ectodomain of membrane-bound proteins, also referred to as ectodomain shedding, has emerged as an important posttranslational regulatory mechanism for modifying the function of cell surface proteins. In line with this notion, several membrane-bound molecules involved in osteoclastogenesis, including CSF-1R and receptor activator of NF-κB ligand (RANKL), are proteolytically cleaved and released from the cell surface. In this study, we investigated whether receptor activator of NF-κB (RANK), one of the most essential molecules in osteoclastogenesis, undergoes ectodomain shedding. The results showed that RANK is released in the form of a soluble monomeric protein and that TNF-α-converting enzyme is involved in this activity. We also identified potential cleavage sites in the juxtamembrane domain of RANK and found that rRANKL induces RANK shedding in a macrophage-like cell line RAW264.7 via TNFR-associated factor 6 and MAPK pathways. Furthermore, we found that RANKL-induced osteoclastogenesis is accelerated in TNF-α-converting enzyme-deficient osteoclast precursors. These observations suggest the potential involvement of ectodomain shedding in the regulation of RANK functions and may provide novel insights into the mechanisms of osteoclastogenesis.
| Original language | English |
|---|---|
| Pages (from-to) | 2442-2448 |
| Number of pages | 7 |
| Journal | Journal of Immunology |
| Volume | 184 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2010 Mar 1 |
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ASJC Scopus subject areas
- Immunology
- Medicine(all)
Cite this
Receptor activator of NF-κB (RANK) ligand induces ectodomain shedding of RANK in murine RAW264.7 macrophages. / Hakozaki, Akihiro; Yoda, Masaki; Tohmonda, Takahide; Furukawa, Mitsuru; Hikata, Tomohiro; Uchikawa, Shinichi; Takaishi, Hironari; Matsumoto, Morio; Chiba, Kazuhiro; Horiuchi, Keisuke; Toyama, Yoshiaki.
In: Journal of Immunology, Vol. 184, No. 5, 01.03.2010, p. 2442-2448.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Receptor activator of NF-κB (RANK) ligand induces ectodomain shedding of RANK in murine RAW264.7 macrophages
AU - Hakozaki, Akihiro
AU - Yoda, Masaki
AU - Tohmonda, Takahide
AU - Furukawa, Mitsuru
AU - Hikata, Tomohiro
AU - Uchikawa, Shinichi
AU - Takaishi, Hironari
AU - Matsumoto, Morio
AU - Chiba, Kazuhiro
AU - Horiuchi, Keisuke
AU - Toyama, Yoshiaki
PY - 2010/3/1
Y1 - 2010/3/1
N2 - Osteoclastogenesis is a highly sophisticated process that involves a variety of membrane-bound proteins expressed in osteoblasts and osteoclast precursors. Over the past several years, proteolytic cleavage and release of the ectodomain of membrane-bound proteins, also referred to as ectodomain shedding, has emerged as an important posttranslational regulatory mechanism for modifying the function of cell surface proteins. In line with this notion, several membrane-bound molecules involved in osteoclastogenesis, including CSF-1R and receptor activator of NF-κB ligand (RANKL), are proteolytically cleaved and released from the cell surface. In this study, we investigated whether receptor activator of NF-κB (RANK), one of the most essential molecules in osteoclastogenesis, undergoes ectodomain shedding. The results showed that RANK is released in the form of a soluble monomeric protein and that TNF-α-converting enzyme is involved in this activity. We also identified potential cleavage sites in the juxtamembrane domain of RANK and found that rRANKL induces RANK shedding in a macrophage-like cell line RAW264.7 via TNFR-associated factor 6 and MAPK pathways. Furthermore, we found that RANKL-induced osteoclastogenesis is accelerated in TNF-α-converting enzyme-deficient osteoclast precursors. These observations suggest the potential involvement of ectodomain shedding in the regulation of RANK functions and may provide novel insights into the mechanisms of osteoclastogenesis.
AB - Osteoclastogenesis is a highly sophisticated process that involves a variety of membrane-bound proteins expressed in osteoblasts and osteoclast precursors. Over the past several years, proteolytic cleavage and release of the ectodomain of membrane-bound proteins, also referred to as ectodomain shedding, has emerged as an important posttranslational regulatory mechanism for modifying the function of cell surface proteins. In line with this notion, several membrane-bound molecules involved in osteoclastogenesis, including CSF-1R and receptor activator of NF-κB ligand (RANKL), are proteolytically cleaved and released from the cell surface. In this study, we investigated whether receptor activator of NF-κB (RANK), one of the most essential molecules in osteoclastogenesis, undergoes ectodomain shedding. The results showed that RANK is released in the form of a soluble monomeric protein and that TNF-α-converting enzyme is involved in this activity. We also identified potential cleavage sites in the juxtamembrane domain of RANK and found that rRANKL induces RANK shedding in a macrophage-like cell line RAW264.7 via TNFR-associated factor 6 and MAPK pathways. Furthermore, we found that RANKL-induced osteoclastogenesis is accelerated in TNF-α-converting enzyme-deficient osteoclast precursors. These observations suggest the potential involvement of ectodomain shedding in the regulation of RANK functions and may provide novel insights into the mechanisms of osteoclastogenesis.
UR - http://www.scopus.com/inward/record.url?scp=77951930123&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77951930123&partnerID=8YFLogxK
U2 - 10.4049/jimmunol.0901188
DO - 10.4049/jimmunol.0901188
M3 - Article
C2 - 20118276
AN - SCOPUS:77951930123
VL - 184
SP - 2442
EP - 2448
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 5
ER -