Cargando…

Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes

Carboxypeptidase Z (CPZ) removes carboxyl-terminal basic amino acid residues, particularly arginine residues, from proteins. CPZ contains a cysteine-rich domain (CRD) similar to the CRD found in the frizzled family of Wnt receptors. We have previously shown that thyroid hormone regulates terminal di...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Lai, Shao, Yvonne Y, Ballock, R Tracy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons and The American Society for Bone and Mineral Research (ASBMR) 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3276606/
https://www.ncbi.nlm.nih.gov/pubmed/18847325
http://dx.doi.org/10.1359/jbmr.081014
_version_ 1782223391964528640
author Wang, Lai
Shao, Yvonne Y
Ballock, R Tracy
author_facet Wang, Lai
Shao, Yvonne Y
Ballock, R Tracy
author_sort Wang, Lai
collection PubMed
description Carboxypeptidase Z (CPZ) removes carboxyl-terminal basic amino acid residues, particularly arginine residues, from proteins. CPZ contains a cysteine-rich domain (CRD) similar to the CRD found in the frizzled family of Wnt receptors. We have previously shown that thyroid hormone regulates terminal differentiation of growth plate chondrocytes through activation of Wnt-4 expression and Wnt/β-catenin signaling. The Wnt-4 protein contains a C-terminal arginine residue and binds to CPZ through the CRD. The objective of this study was to determine whether CPZ modulates Wnt/β-catenin signaling and terminal differentiation of growth plate chondrocytes. Our results show that CPZ and Wnt-4 mRNA are co-expressed throughout growth plate cartilage. In primary pellet cultures of rat growth plate chondrocytes, thyroid hormone increases both Wnt-4 and CPZ expression, as well as CPZ enzymatic activity. Knockdown of either Wnt-4 or CPZ mRNA levels using an RNA interference technique or blocking CPZ enzymatic activity with the carboxypeptidase inhibitor GEMSA reduces the thyroid hormone effect on both alkaline phosphatase activity and Col10a1 mRNA expression. Adenoviral overexpression of CPZ activates Wnt/β-catenin signaling and promotes the terminal differentiation of growth plate cells. Overexpression of CPZ in growth plate chondrocytes also removes the C-terminal arginine residue from a synthetic peptide consisting of the carboxyl-terminal 16 amino acids of the Wnt-4 protein. Removal of the C-terminal arginine residue of Wnt-4 by site-directed mutagenesis enhances the positive effect of Wnt-4 on terminal differentiation. These data indicate that thyroid hormone may regulate terminal differentiation of growth plate chondrocytes in part by modulating Wnt signaling pathways through the induction of CPZ and subsequent CPZ-enhanced activation of Wnt-4.
format Online
Article
Text
id pubmed-3276606
institution National Center for Biotechnology Information
language English
publishDate 2009
publisher John Wiley and Sons and The American Society for Bone and Mineral Research (ASBMR)
record_format MEDLINE/PubMed
spelling pubmed-32766062012-02-13 Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes Wang, Lai Shao, Yvonne Y Ballock, R Tracy J Bone Miner Res Research-Articles Carboxypeptidase Z (CPZ) removes carboxyl-terminal basic amino acid residues, particularly arginine residues, from proteins. CPZ contains a cysteine-rich domain (CRD) similar to the CRD found in the frizzled family of Wnt receptors. We have previously shown that thyroid hormone regulates terminal differentiation of growth plate chondrocytes through activation of Wnt-4 expression and Wnt/β-catenin signaling. The Wnt-4 protein contains a C-terminal arginine residue and binds to CPZ through the CRD. The objective of this study was to determine whether CPZ modulates Wnt/β-catenin signaling and terminal differentiation of growth plate chondrocytes. Our results show that CPZ and Wnt-4 mRNA are co-expressed throughout growth plate cartilage. In primary pellet cultures of rat growth plate chondrocytes, thyroid hormone increases both Wnt-4 and CPZ expression, as well as CPZ enzymatic activity. Knockdown of either Wnt-4 or CPZ mRNA levels using an RNA interference technique or blocking CPZ enzymatic activity with the carboxypeptidase inhibitor GEMSA reduces the thyroid hormone effect on both alkaline phosphatase activity and Col10a1 mRNA expression. Adenoviral overexpression of CPZ activates Wnt/β-catenin signaling and promotes the terminal differentiation of growth plate cells. Overexpression of CPZ in growth plate chondrocytes also removes the C-terminal arginine residue from a synthetic peptide consisting of the carboxyl-terminal 16 amino acids of the Wnt-4 protein. Removal of the C-terminal arginine residue of Wnt-4 by site-directed mutagenesis enhances the positive effect of Wnt-4 on terminal differentiation. These data indicate that thyroid hormone may regulate terminal differentiation of growth plate chondrocytes in part by modulating Wnt signaling pathways through the induction of CPZ and subsequent CPZ-enhanced activation of Wnt-4. John Wiley and Sons and The American Society for Bone and Mineral Research (ASBMR) 2009-02 2008-10-13 /pmc/articles/PMC3276606/ /pubmed/18847325 http://dx.doi.org/10.1359/jbmr.081014 Text en Copyright © 2009 American Society for Bone and Mineral Research http://creativecommons.org/licenses/by/2.5/ Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
spellingShingle Research-Articles
Wang, Lai
Shao, Yvonne Y
Ballock, R Tracy
Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes
title Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes
title_full Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes
title_fullStr Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes
title_full_unstemmed Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes
title_short Carboxypeptidase Z (CPZ) Links Thyroid Hormone and Wnt Signaling Pathways in Growth Plate Chondrocytes
title_sort carboxypeptidase z (cpz) links thyroid hormone and wnt signaling pathways in growth plate chondrocytes
topic Research-Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3276606/
https://www.ncbi.nlm.nih.gov/pubmed/18847325
http://dx.doi.org/10.1359/jbmr.081014
work_keys_str_mv AT wanglai carboxypeptidasezcpzlinksthyroidhormoneandwntsignalingpathwaysingrowthplatechondrocytes
AT shaoyvonney carboxypeptidasezcpzlinksthyroidhormoneandwntsignalingpathwaysingrowthplatechondrocytes
AT ballockrtracy carboxypeptidasezcpzlinksthyroidhormoneandwntsignalingpathwaysingrowthplatechondrocytes