title: Characterization of a non-fibrillar-related collagen in the mollusc Haliotis tuberculata and its biological activity on human dermal fibroblasts.
abstract: In invertebrates, members of the collagen family have been found in various phyla. Surprisingly, in mollusc, little is known about such molecules. In this study, we characterize the full-length abalone type IV collagen and we analysed its biological effects on human fibroblast in order to gain insights about this molecule in molluscs and particularly clues about its roles. We screened a cDNA library of Haliotis tuberculata hemocytes. The expression pattern of the transcript is determined using real-time polymerase chain reaction and in situ hybridization. The close identity between α1(IV) C-terminal domain and the vertebrate homologue led us to produce, purify and test in vitro a recombinant protein corresponding to this region using human dermal fibroblasts cell culture. The biological effects were evaluated on proliferation and on differentiation. We found that the 5,334-bp open reading frame transcript encodes a protein of 1,777 amino acids, including an interrupted 1,502-residue collagenous domain and a 232-residue C-terminal non-collagenous domain. The expression pattern of this transcript is mainly found in the mantle and hemocytes. The recombinant protein corresponding α1(IV) C-terminal domain increased fibroblast proliferation by 69% and doubled collagen synthesis produced in primary cultures. This work provides the first complete primary structure of a mollusc non-fibrillar collagen chain and the biological effects of its C-terminal domain on human cells. In this study, we prove that the NC1 domain from a molluscan collagen can improve human fibroblast proliferation as well as differentiation.
authors.affiliation: UMR 100 IFREMER Physiologie et Ecophysiologie des Mollusques Marins-IFR 146 ICORE-IBFA-Esplanade de la Paix, Université de Caen Basse-normandie, 14032, Caen, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Fleury
authors.forename: Antoine
authors.initials: A
authors.lastname: Serpentini
authors.forename: Magdalini
authors.initials: M
authors.lastname: Kypriotou
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Renard
authors.forename: Philippe
authors.initials: P
authors.lastname: Galéra
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Lebel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D008974:Mollusca / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D003094:Collagen