[en] Mutable collagenous tissues (MCTs) from echinoderms (e.g., sea stars, sea urchins) possess the remarkable ability to change their mechanical properties rapidly and reversibly thanks to the release of effector molecules regulating the number of cross-links between collagen fibrils. Among these effector molecules, tensilin has been identified as a stiffening factor in sea cucumber MCTs. Since its discovery and description twenty years ago, tensilin orthologs have been identified in a few sea cucumber species but no novel information about its molecular mode of action has been reported. In this study, using a combination of in silico analyses, we identified the tensilin present in the dermis of Holothuria forskali, Hf-(D)Tensilin. Anti-peptide antibodies showed that this protein is localised in the secretory granules of type 2 juxtaligamental-like cells, a MCT specific cell type. We then used the bacterium E. coli to produce recombinantly Hf-(D)Tensilin and confirmed its stiffening effect on pieces of the dermis and its aggregation effect on collagen fibrils extracted from the sea cucumber dermis. To investigate how tensilin can cross-bridge collagen fibrils, truncated recombinant tensilins were also produced and used in combination with various compounds. Results suggest that two types of interactions contribute to the aggregation effect of tensilin on the fibrils: (1) the N-terminal NTR TIMP like domain of the protein interacts strongly with sulfated GAGs attached to the surface of the collagen fibrils, and (2) the C-terminal part of the protein is involved in its dimerisation/oligomerisation through ionic but possibly also cation-π and hydrophobic interactions.
Bonneel, Marie ✱; Biology of Marine Organisms and Biomimetics Unit, Research Institute for Biosciences, University of Mons, Place du Parc 23, B-7000 Mons, Belgium
Hennebert, Elise ✱; Université de Mons - UMONS > Faculté des Sciences > Service de Biologie cellulaire
Aranko, A Sesilja ; Department of Bioproducts and Biosystems, School of Chemical Engineering, Aalto University, P.O. Box 16100, FI-02150 Espoo, Finland
Hwang, Dong Soo; Division of Environmental Science and Engineering, Pohang University of Science and Technology, Pohang 37673, Republic of Korea
Lefevre, Mathilde; Laboratory of Cell Biology, Research Institute for Biosciences, University of Mons, Place du Parc 23, B-7000 Mons, Belgium
Pommier, Valentine; Biology of Marine Organisms and Biomimetics Unit, Research Institute for Biosciences, University of Mons, Place du Parc 23, B-7000 Mons, Belgium
Wattiez, Ruddy ; Université de Mons - UMONS > Faculté des Sciences > Service de Protéomie et Microbiologie
Delroisse, Jérôme ; Biology of Marine Organisms and Biomimetics Unit, Research Institute for Biosciences, University of Mons, Place du Parc 23, B-7000 Mons, Belgium
Flammang, Patrick ; Université de Mons - UMONS > Faculté des Sciences > Service de Biologie des Organismes Marins et Biomimétisme
✱ These authors have contributed equally to this work.
Language :
English
Title :
Molecular mechanisms mediating stiffening in the mechanically adaptable connective tissues of sea cucumbers.
This work was supported by the Fund for Scientific Research of Belgium (F.R.S.-FNRS) FRIA doctoral grant 17953 (M.B.); the Fund for Medical Research in Hainaut (F.R.M.H.) (E.H.); the Academy of Finland projects 308772, 333238 (A.S.A.); the Academy of Finland's Center of Excellence programme 20142019 (A.S.A.); the Fédération Wallonie-Bruxelles – Actions de Recherche Concertées ARC project “PROTEST”, ARC-17/21 UMONS 3 (E.H., R.W., P.F.); and the Fund for Scientific Research of Belgium (F.R.S.-FNRS) “Projet de Recherche” T.0088.20 (P.F.)The authors thank Nathan Puozzo who designed the schematic model in Fig. 8 . The bioprofiling platform used for the de novo peptide sequencing analysis was supported by the European Regional Development Fund and the Walloon Region. J.D. and P.F. are, respectively, Postdoctoral Researcher and Research Director of the F.R.S.-FNRS. This study is a contribution of the Centre Interuniversitaire de Biologie Marine.The authors thank Nathan Puozzo who designed the schematic model in Fig. 8. The bioprofiling platform used for the de novo peptide sequencing analysis was supported by the European Regional Development Fund and the Walloon Region. J.D. and P.F. are, respectively, Postdoctoral Researcher and Research Director of the F.R.S.-FNRS. This study is a contribution of the Centre Interuniversitaire de Biologie Marine. This work was supported by the Fund for Scientific Research of Belgium (F.R.S.-FNRS) FRIA doctoral grant 17953 (M.B.); the Fund for Medical Research in Hainaut (F.R.M.H.) (E.H.); the Academy of Finland projects 308772, 333238 (A.S.A.); the Academy of Finland's Center of Excellence programme 20142019 (A.S.A.); the F?d?ration Wallonie-Bruxelles ? Actions de Recherche Concert?es ARC project ?PROTEST?, ARC-17/21 UMONS 3 (E.H. R.W. P.F.); and the Fund for Scientific Research of Belgium (F.R.S.-FNRS) ?Projet de Recherche? T.0088.20 (P.F.), M.B. E.H. and P.F. designed research; M.B. E.H. M.L. V.P. J.D. R.W. and P.F. performed experiments; M.B. E.H. A.S.A. J.D. D.S.H. and P.F. analysed data; M.B. E.H. J.D. and P.F wrote the first draft of the paper. All co-authors participated in discussions and revised the final manuscript. All data are available in the main text or the supplementary materials.
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