Cargando…

The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2

Cellular Communication Network (CCN) proteins have multimodular structures important for their roles in cellular responses associated with organ development and tissue homeostasis. CCN2 has previously been reported to be secreted as a preproprotein that requires proteolytic activation to release its...

Descripción completa

Detalles Bibliográficos
Autores principales: Zolfaghari, Sima, Kaasbøll, Ole Jørgen, Monsen, Vivi T., Sredic, Bojana, Hagelin, Else Marie V., Attramadal, Håvard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9860493/
https://www.ncbi.nlm.nih.gov/pubmed/36529291
http://dx.doi.org/10.1016/j.jbc.2022.102803
_version_ 1784874595647488000
author Zolfaghari, Sima
Kaasbøll, Ole Jørgen
Monsen, Vivi T.
Sredic, Bojana
Hagelin, Else Marie V.
Attramadal, Håvard
author_facet Zolfaghari, Sima
Kaasbøll, Ole Jørgen
Monsen, Vivi T.
Sredic, Bojana
Hagelin, Else Marie V.
Attramadal, Håvard
author_sort Zolfaghari, Sima
collection PubMed
description Cellular Communication Network (CCN) proteins have multimodular structures important for their roles in cellular responses associated with organ development and tissue homeostasis. CCN2 has previously been reported to be secreted as a preproprotein that requires proteolytic activation to release its bioactive carboxyl-terminal fragment. Here, our goal was to resolve whether CCN5, a divergent member of the CCN family with converse functions relative to CCN2, releases the TSP1 homology domain as its bioactive signaling entity. The recombinant CCN5 or CCN3 TSP1 homology domains were produced in ExpiCHO-S or DG44 CHO cells as secretory fusion proteins appended to the carboxyl-terminal end of His-Halo-Sumo or amino-terminal end of human albumin and purified from the cell culture medium. We tested these fusion proteins in various phosphokinase signaling pathways or cell physiologic assays. Fusion proteins with the CCN5 TSP1 domain inhibited key signaling pathways previously reported to be stimulated by CCN2, irrespective of fusion partner. The fusion proteins also efficiently inhibited CCN1/2-stimulated cell migration and gap closure following scratch wound of fibroblasts. Fusion protein with the CCN3 TSP1 domain inhibited these functions with similar efficacy and potency as that of the CCN5 TSP1 domain. The CCN5 TSP1 domain also recapitulated a positive regulatory function previously assigned to full-length CCN5, that is, induction of estrogen receptor-α mRNA expression in triple negative MDA-MB-231 mammary adenocarcinoma cells and inhibited epithelial-to-mesenchymal transition and CCN2-induced mammosphere formation of MCF-7 adenocarcinoma cells. In conclusion, the CCN5 TSP1 domain is the bioactive entity that confers the biologic functions of unprocessed CCN5.
format Online
Article
Text
id pubmed-9860493
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher American Society for Biochemistry and Molecular Biology
record_format MEDLINE/PubMed
spelling pubmed-98604932023-01-26 The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2 Zolfaghari, Sima Kaasbøll, Ole Jørgen Monsen, Vivi T. Sredic, Bojana Hagelin, Else Marie V. Attramadal, Håvard J Biol Chem Research Article Cellular Communication Network (CCN) proteins have multimodular structures important for their roles in cellular responses associated with organ development and tissue homeostasis. CCN2 has previously been reported to be secreted as a preproprotein that requires proteolytic activation to release its bioactive carboxyl-terminal fragment. Here, our goal was to resolve whether CCN5, a divergent member of the CCN family with converse functions relative to CCN2, releases the TSP1 homology domain as its bioactive signaling entity. The recombinant CCN5 or CCN3 TSP1 homology domains were produced in ExpiCHO-S or DG44 CHO cells as secretory fusion proteins appended to the carboxyl-terminal end of His-Halo-Sumo or amino-terminal end of human albumin and purified from the cell culture medium. We tested these fusion proteins in various phosphokinase signaling pathways or cell physiologic assays. Fusion proteins with the CCN5 TSP1 domain inhibited key signaling pathways previously reported to be stimulated by CCN2, irrespective of fusion partner. The fusion proteins also efficiently inhibited CCN1/2-stimulated cell migration and gap closure following scratch wound of fibroblasts. Fusion protein with the CCN3 TSP1 domain inhibited these functions with similar efficacy and potency as that of the CCN5 TSP1 domain. The CCN5 TSP1 domain also recapitulated a positive regulatory function previously assigned to full-length CCN5, that is, induction of estrogen receptor-α mRNA expression in triple negative MDA-MB-231 mammary adenocarcinoma cells and inhibited epithelial-to-mesenchymal transition and CCN2-induced mammosphere formation of MCF-7 adenocarcinoma cells. In conclusion, the CCN5 TSP1 domain is the bioactive entity that confers the biologic functions of unprocessed CCN5. American Society for Biochemistry and Molecular Biology 2022-12-15 /pmc/articles/PMC9860493/ /pubmed/36529291 http://dx.doi.org/10.1016/j.jbc.2022.102803 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Zolfaghari, Sima
Kaasbøll, Ole Jørgen
Monsen, Vivi T.
Sredic, Bojana
Hagelin, Else Marie V.
Attramadal, Håvard
The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2
title The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2
title_full The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2
title_fullStr The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2
title_full_unstemmed The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2
title_short The carboxyl-terminal TSP1-homology domain is the biologically active effector peptide of matricellular protein CCN5 that counteracts profibrotic CCN2
title_sort carboxyl-terminal tsp1-homology domain is the biologically active effector peptide of matricellular protein ccn5 that counteracts profibrotic ccn2
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9860493/
https://www.ncbi.nlm.nih.gov/pubmed/36529291
http://dx.doi.org/10.1016/j.jbc.2022.102803
work_keys_str_mv AT zolfagharisima thecarboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT kaasbøllolejørgen thecarboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT monsenvivit thecarboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT sredicbojana thecarboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT hagelinelsemariev thecarboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT attramadalhavard thecarboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT zolfagharisima carboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT kaasbøllolejørgen carboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT monsenvivit carboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT sredicbojana carboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT hagelinelsemariev carboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2
AT attramadalhavard carboxylterminaltsp1homologydomainisthebiologicallyactiveeffectorpeptideofmatricellularproteinccn5thatcounteractsprofibroticccn2