Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis

Jinmoon Kim, Sungil Jang, Aeryun Kim, Hanfu Su, Niluka Gunawardhana, Yeong Eui Jeon, Eun Jung Bak, Ji Hye Kim, JeongHeon Cha

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Mammalian γ-glutamyltranspeptidase (GGT) has been identified as a bone-resorbing factor. Since GGT of Bacillus subtilis exhibits similarity in their primary structure and enzymatic characteristics with mammalian GGTs, the bone-resorbing activity of bacterial GGT was examined in this study. Osteoclastogenesis was performed in a co-culture system of mouse calvaria-derived osteoblasts and bone marrow cells. A conditioned medium from GGT-overproducing B. subtilis culture showed significantly higher activity of osteoclast formation than a conditioned medium from wild-type B. subtilis culture. Recombinant GGT (rGGT) of wild-type B. subtilis and an enzymatic activity-defected rGGT of B. subtilis 2288 mutant were expressed in Escherichia coli and purified using His tag. Both purified rGGTs induced similar levels of osteoclastogenesis, suggesting that B. subtilis GGT possesses virulent boneresorbing activity and its activity is probably independent of its enzymatic activity. Furthermore, a recombinant protein of B. subtilis GGT heavy subunit (Bs rGGT/H) showed strong activity of osteoclastogenesis while the light subunit failed to show strong activity, suggesting that the bone-resorbing activity is mainly located at the heavy subunit. More importantly, the GGT enzymatic activity may not be required for this virulence activity since the light subunit contains the catalytic pocket. In addition, B. subtilis rGGT stimulated mRNA expressions of receptor activator of nuclear factor kappa-B ligand (RANKL) and cyclooxygenase-2 (COX-2), while an osteoprotegerin inhibited the osteoclast formation induced by Bs rGGT/H. This is the first demonstration that bacterial GGT itself is sufficient to act as a bone-resorbing virulence factor via RANKL-dependent pathway. Therefore, it can be hypothesized that GGT of periodontopathic bacteria may play an important role as a virulence factor in bone destruction.

Original languageEnglish
Pages (from-to)396-402
Number of pages7
JournalJournal of Microbiology
Volume54
Issue number5
DOIs
Publication statusPublished - 2016 Jan 1

Fingerprint

Virulence Factors
Bacillus subtilis
Bone and Bones
Osteogenesis
RANK Ligand
Osteoclasts
Conditioned Culture Medium
Light
Osteoprotegerin
Cyclooxygenase 2
Coculture Techniques
Osteoblasts
Recombinant Proteins
Skull
Bone Marrow Cells
Virulence
Catalytic Domain
Escherichia coli
Bacteria
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Applied Microbiology and Biotechnology

Cite this

Kim, Jinmoon ; Jang, Sungil ; Kim, Aeryun ; Su, Hanfu ; Gunawardhana, Niluka ; Jeon, Yeong Eui ; Bak, Eun Jung ; Kim, Ji Hye ; Cha, JeongHeon. / Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis. In: Journal of Microbiology. 2016 ; Vol. 54, No. 5. pp. 396-402.
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title = "Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis",
abstract = "Mammalian γ-glutamyltranspeptidase (GGT) has been identified as a bone-resorbing factor. Since GGT of Bacillus subtilis exhibits similarity in their primary structure and enzymatic characteristics with mammalian GGTs, the bone-resorbing activity of bacterial GGT was examined in this study. Osteoclastogenesis was performed in a co-culture system of mouse calvaria-derived osteoblasts and bone marrow cells. A conditioned medium from GGT-overproducing B. subtilis culture showed significantly higher activity of osteoclast formation than a conditioned medium from wild-type B. subtilis culture. Recombinant GGT (rGGT) of wild-type B. subtilis and an enzymatic activity-defected rGGT of B. subtilis 2288 mutant were expressed in Escherichia coli and purified using His tag. Both purified rGGTs induced similar levels of osteoclastogenesis, suggesting that B. subtilis GGT possesses virulent boneresorbing activity and its activity is probably independent of its enzymatic activity. Furthermore, a recombinant protein of B. subtilis GGT heavy subunit (Bs rGGT/H) showed strong activity of osteoclastogenesis while the light subunit failed to show strong activity, suggesting that the bone-resorbing activity is mainly located at the heavy subunit. More importantly, the GGT enzymatic activity may not be required for this virulence activity since the light subunit contains the catalytic pocket. In addition, B. subtilis rGGT stimulated mRNA expressions of receptor activator of nuclear factor kappa-B ligand (RANKL) and cyclooxygenase-2 (COX-2), while an osteoprotegerin inhibited the osteoclast formation induced by Bs rGGT/H. This is the first demonstration that bacterial GGT itself is sufficient to act as a bone-resorbing virulence factor via RANKL-dependent pathway. Therefore, it can be hypothesized that GGT of periodontopathic bacteria may play an important role as a virulence factor in bone destruction.",
author = "Jinmoon Kim and Sungil Jang and Aeryun Kim and Hanfu Su and Niluka Gunawardhana and Jeon, {Yeong Eui} and Bak, {Eun Jung} and Kim, {Ji Hye} and JeongHeon Cha",
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Kim, J, Jang, S, Kim, A, Su, H, Gunawardhana, N, Jeon, YE, Bak, EJ, Kim, JH & Cha, J 2016, 'Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis', Journal of Microbiology, vol. 54, no. 5, pp. 396-402. https://doi.org/10.1007/s12275-016-6137-1

Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis. / Kim, Jinmoon; Jang, Sungil; Kim, Aeryun; Su, Hanfu; Gunawardhana, Niluka; Jeon, Yeong Eui; Bak, Eun Jung; Kim, Ji Hye; Cha, JeongHeon.

In: Journal of Microbiology, Vol. 54, No. 5, 01.01.2016, p. 396-402.

Research output: Contribution to journalArticle

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T1 - Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis

AU - Kim, Jinmoon

AU - Jang, Sungil

AU - Kim, Aeryun

AU - Su, Hanfu

AU - Gunawardhana, Niluka

AU - Jeon, Yeong Eui

AU - Bak, Eun Jung

AU - Kim, Ji Hye

AU - Cha, JeongHeon

PY - 2016/1/1

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N2 - Mammalian γ-glutamyltranspeptidase (GGT) has been identified as a bone-resorbing factor. Since GGT of Bacillus subtilis exhibits similarity in their primary structure and enzymatic characteristics with mammalian GGTs, the bone-resorbing activity of bacterial GGT was examined in this study. Osteoclastogenesis was performed in a co-culture system of mouse calvaria-derived osteoblasts and bone marrow cells. A conditioned medium from GGT-overproducing B. subtilis culture showed significantly higher activity of osteoclast formation than a conditioned medium from wild-type B. subtilis culture. Recombinant GGT (rGGT) of wild-type B. subtilis and an enzymatic activity-defected rGGT of B. subtilis 2288 mutant were expressed in Escherichia coli and purified using His tag. Both purified rGGTs induced similar levels of osteoclastogenesis, suggesting that B. subtilis GGT possesses virulent boneresorbing activity and its activity is probably independent of its enzymatic activity. Furthermore, a recombinant protein of B. subtilis GGT heavy subunit (Bs rGGT/H) showed strong activity of osteoclastogenesis while the light subunit failed to show strong activity, suggesting that the bone-resorbing activity is mainly located at the heavy subunit. More importantly, the GGT enzymatic activity may not be required for this virulence activity since the light subunit contains the catalytic pocket. In addition, B. subtilis rGGT stimulated mRNA expressions of receptor activator of nuclear factor kappa-B ligand (RANKL) and cyclooxygenase-2 (COX-2), while an osteoprotegerin inhibited the osteoclast formation induced by Bs rGGT/H. This is the first demonstration that bacterial GGT itself is sufficient to act as a bone-resorbing virulence factor via RANKL-dependent pathway. Therefore, it can be hypothesized that GGT of periodontopathic bacteria may play an important role as a virulence factor in bone destruction.

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