Identification of human asparaginyl endopeptidase (legumain) as an inhibitor of osteoclast formation and bone resorption

Sun Jin Choi, Sakamuri V. Reddy, Rowena D. Devlin, Cheikh Menaa, Hoyeon Chung, Brendan F. Boyce, G. David Roodman

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

We screened a human osteoclast (OCL) cDNA expression library for OCL inhibitory factors and identified a clone that blocked both human and murine OCL formation and bone resorption by more than 60%. This clone was identical to human legumain, a cysteine endopeptidase. Legumain significantly inhibited OCL-like multinucleated cell formation induced by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and parathyroid hormone-related protein (PTHrP) in mouse and human bone marrow cultures, and bone resorption in the fetal rat long bone assay in a dose-dependent manner. Legumain was detected in freshly isolated marrow plasma from normal donors and conditioned media from human marrow cultures. Furthermore, treatment of human marrow cultures with an antibody to legumain induced OCL formation to levels that were as high as those induced by 1,25-(OH)2D3. Implantation in nude mice of 293 cells transfected with the legumain cDNA and constitutively expressing high levels of the protein significantly reduced hypercalcemia induced by PTHrP by about 50%, and significantly inhibited the increase in OCL surface and in OCL number expressed per mm2 bone area and per mm bone surface induced by PTHrP. These results suggest that legumain may be a physiologic local regulator of OCL activity that can negatively modulate OCL formation and activity.

Original languageEnglish (US)
Pages (from-to)27747-27753
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number39
DOIs
StatePublished - Sep 24 1999
Externally publishedYes

Fingerprint

asparaginylendopeptidase
Forensic Anthropology
Osteoclasts
Bone Resorption
Bone
Parathyroid Hormone-Related Protein
Bone Marrow
Bone and Bones
Cysteine Endopeptidases
Complementary DNA
Clone Cells
Calcitriol
Hypercalcemia
Conditioned Culture Medium

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of human asparaginyl endopeptidase (legumain) as an inhibitor of osteoclast formation and bone resorption. / Choi, Sun Jin; Reddy, Sakamuri V.; Devlin, Rowena D.; Menaa, Cheikh; Chung, Hoyeon; Boyce, Brendan F.; Roodman, G. David.

In: Journal of Biological Chemistry, Vol. 274, No. 39, 24.09.1999, p. 27747-27753.

Research output: Contribution to journalArticle

Choi, Sun Jin ; Reddy, Sakamuri V. ; Devlin, Rowena D. ; Menaa, Cheikh ; Chung, Hoyeon ; Boyce, Brendan F. ; Roodman, G. David. / Identification of human asparaginyl endopeptidase (legumain) as an inhibitor of osteoclast formation and bone resorption. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 39. pp. 27747-27753.
@article{f3d4ed5f6ad94cb3a8f5aae86f486b7d,
title = "Identification of human asparaginyl endopeptidase (legumain) as an inhibitor of osteoclast formation and bone resorption",
abstract = "We screened a human osteoclast (OCL) cDNA expression library for OCL inhibitory factors and identified a clone that blocked both human and murine OCL formation and bone resorption by more than 60{\%}. This clone was identical to human legumain, a cysteine endopeptidase. Legumain significantly inhibited OCL-like multinucleated cell formation induced by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and parathyroid hormone-related protein (PTHrP) in mouse and human bone marrow cultures, and bone resorption in the fetal rat long bone assay in a dose-dependent manner. Legumain was detected in freshly isolated marrow plasma from normal donors and conditioned media from human marrow cultures. Furthermore, treatment of human marrow cultures with an antibody to legumain induced OCL formation to levels that were as high as those induced by 1,25-(OH)2D3. Implantation in nude mice of 293 cells transfected with the legumain cDNA and constitutively expressing high levels of the protein significantly reduced hypercalcemia induced by PTHrP by about 50{\%}, and significantly inhibited the increase in OCL surface and in OCL number expressed per mm2 bone area and per mm bone surface induced by PTHrP. These results suggest that legumain may be a physiologic local regulator of OCL activity that can negatively modulate OCL formation and activity.",
author = "Choi, {Sun Jin} and Reddy, {Sakamuri V.} and Devlin, {Rowena D.} and Cheikh Menaa and Hoyeon Chung and Boyce, {Brendan F.} and Roodman, {G. David}",
year = "1999",
month = "9",
day = "24",
doi = "10.1074/jbc.274.39.27747",
language = "English (US)",
volume = "274",
pages = "27747--27753",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "39",

}

TY - JOUR

T1 - Identification of human asparaginyl endopeptidase (legumain) as an inhibitor of osteoclast formation and bone resorption

AU - Choi, Sun Jin

AU - Reddy, Sakamuri V.

AU - Devlin, Rowena D.

AU - Menaa, Cheikh

AU - Chung, Hoyeon

AU - Boyce, Brendan F.

AU - Roodman, G. David

PY - 1999/9/24

Y1 - 1999/9/24

N2 - We screened a human osteoclast (OCL) cDNA expression library for OCL inhibitory factors and identified a clone that blocked both human and murine OCL formation and bone resorption by more than 60%. This clone was identical to human legumain, a cysteine endopeptidase. Legumain significantly inhibited OCL-like multinucleated cell formation induced by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and parathyroid hormone-related protein (PTHrP) in mouse and human bone marrow cultures, and bone resorption in the fetal rat long bone assay in a dose-dependent manner. Legumain was detected in freshly isolated marrow plasma from normal donors and conditioned media from human marrow cultures. Furthermore, treatment of human marrow cultures with an antibody to legumain induced OCL formation to levels that were as high as those induced by 1,25-(OH)2D3. Implantation in nude mice of 293 cells transfected with the legumain cDNA and constitutively expressing high levels of the protein significantly reduced hypercalcemia induced by PTHrP by about 50%, and significantly inhibited the increase in OCL surface and in OCL number expressed per mm2 bone area and per mm bone surface induced by PTHrP. These results suggest that legumain may be a physiologic local regulator of OCL activity that can negatively modulate OCL formation and activity.

AB - We screened a human osteoclast (OCL) cDNA expression library for OCL inhibitory factors and identified a clone that blocked both human and murine OCL formation and bone resorption by more than 60%. This clone was identical to human legumain, a cysteine endopeptidase. Legumain significantly inhibited OCL-like multinucleated cell formation induced by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and parathyroid hormone-related protein (PTHrP) in mouse and human bone marrow cultures, and bone resorption in the fetal rat long bone assay in a dose-dependent manner. Legumain was detected in freshly isolated marrow plasma from normal donors and conditioned media from human marrow cultures. Furthermore, treatment of human marrow cultures with an antibody to legumain induced OCL formation to levels that were as high as those induced by 1,25-(OH)2D3. Implantation in nude mice of 293 cells transfected with the legumain cDNA and constitutively expressing high levels of the protein significantly reduced hypercalcemia induced by PTHrP by about 50%, and significantly inhibited the increase in OCL surface and in OCL number expressed per mm2 bone area and per mm bone surface induced by PTHrP. These results suggest that legumain may be a physiologic local regulator of OCL activity that can negatively modulate OCL formation and activity.

UR - http://www.scopus.com/inward/record.url?scp=0001460158&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0001460158&partnerID=8YFLogxK

U2 - 10.1074/jbc.274.39.27747

DO - 10.1074/jbc.274.39.27747

M3 - Article

C2 - 10488118

AN - SCOPUS:0001460158

VL - 274

SP - 27747

EP - 27753

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 39

ER -