Cargando…

The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration

Testicans are modular proteoglycans of the extracellular matrix of various tissues where they contribute to matrix integrity and exert cellular effects like neurite outgrowth and cell migration. Using testican-2 as a representative member of the family, we tackle the complete lack of general structu...

Descripción completa

Detalles Bibliográficos
Autores principales: Krajnc, Anja, Gaber, Aljaž, Lenarčič, Brigita, Pavšič, Miha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7763218/
https://www.ncbi.nlm.nih.gov/pubmed/33321927
http://dx.doi.org/10.3390/ijms21249413
_version_ 1783627965809033216
author Krajnc, Anja
Gaber, Aljaž
Lenarčič, Brigita
Pavšič, Miha
author_facet Krajnc, Anja
Gaber, Aljaž
Lenarčič, Brigita
Pavšič, Miha
author_sort Krajnc, Anja
collection PubMed
description Testicans are modular proteoglycans of the extracellular matrix of various tissues where they contribute to matrix integrity and exert cellular effects like neurite outgrowth and cell migration. Using testican-2 as a representative member of the family, we tackle the complete lack of general structural information and structure–function relationship. First, we show using isothermal titration calorimetry and modeling that extracellular calcium-binding domain (EC) has only one active calcium-binding site, while the other potential site is inactive, and that testican-2 is within extracellular matrix always in the calcium-loaded form. Next, we demonstrate using various prediction methods that N- and C-terminal regions plus interdomain connections are flexible. We support this by small-angle X-ray-scattering analysis of C-terminally truncated testican-2, which indicates that the triplet follistatin-EC-thyroglobulin domain forms a moderately compact core while the unique N-terminal is disordered. Finally, using cell exclusion zone assay, we show that it is this domain triplet that is responsible for promoting cell migration and not the N- and C-terminal regions.
format Online
Article
Text
id pubmed-7763218
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-77632182020-12-27 The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration Krajnc, Anja Gaber, Aljaž Lenarčič, Brigita Pavšič, Miha Int J Mol Sci Article Testicans are modular proteoglycans of the extracellular matrix of various tissues where they contribute to matrix integrity and exert cellular effects like neurite outgrowth and cell migration. Using testican-2 as a representative member of the family, we tackle the complete lack of general structural information and structure–function relationship. First, we show using isothermal titration calorimetry and modeling that extracellular calcium-binding domain (EC) has only one active calcium-binding site, while the other potential site is inactive, and that testican-2 is within extracellular matrix always in the calcium-loaded form. Next, we demonstrate using various prediction methods that N- and C-terminal regions plus interdomain connections are flexible. We support this by small-angle X-ray-scattering analysis of C-terminally truncated testican-2, which indicates that the triplet follistatin-EC-thyroglobulin domain forms a moderately compact core while the unique N-terminal is disordered. Finally, using cell exclusion zone assay, we show that it is this domain triplet that is responsible for promoting cell migration and not the N- and C-terminal regions. MDPI 2020-12-10 /pmc/articles/PMC7763218/ /pubmed/33321927 http://dx.doi.org/10.3390/ijms21249413 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Krajnc, Anja
Gaber, Aljaž
Lenarčič, Brigita
Pavšič, Miha
The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration
title The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration
title_full The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration
title_fullStr The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration
title_full_unstemmed The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration
title_short The Central Region of Testican-2 Forms a Compact Core and Promotes Cell Migration
title_sort central region of testican-2 forms a compact core and promotes cell migration
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7763218/
https://www.ncbi.nlm.nih.gov/pubmed/33321927
http://dx.doi.org/10.3390/ijms21249413
work_keys_str_mv AT krajncanja thecentralregionoftestican2formsacompactcoreandpromotescellmigration
AT gaberaljaz thecentralregionoftestican2formsacompactcoreandpromotescellmigration
AT lenarcicbrigita thecentralregionoftestican2formsacompactcoreandpromotescellmigration
AT pavsicmiha thecentralregionoftestican2formsacompactcoreandpromotescellmigration
AT krajncanja centralregionoftestican2formsacompactcoreandpromotescellmigration
AT gaberaljaz centralregionoftestican2formsacompactcoreandpromotescellmigration
AT lenarcicbrigita centralregionoftestican2formsacompactcoreandpromotescellmigration
AT pavsicmiha centralregionoftestican2formsacompactcoreandpromotescellmigration