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.
Mar Biotechnol (NY). 2011 Oct;13(5):1003-16. doi: 10.1007/s10126-011-9364-9. Epub 2011 Jan 27.
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.
在无脊椎动物中，已经在各种门中发现了胶原蛋白家族的成员。令人惊讶的是，在软体动物中，对这类分子知之甚少。在这项研究中，我们对完整的鲍鱼 IV 型胶原蛋白进行了特征描述，并分析了其对人成纤维细胞的生物学效应，以便深入了解软体动物中的这种分子，特别是其作用的线索。我们筛选了鲍血细胞的 cDNA 文库。使用实时聚合酶链反应和原位杂交确定了转录本的表达模式。α1(IV)C 端结构域与脊椎动物同源物之间的高度同源性使我们能够使用人真皮成纤维细胞培养物生产、纯化和测试对应于该区域的重组蛋白。评估了生物效应在增殖和分化上的影响。我们发现，5334bp 的开放阅读框转录本编码 1777 个氨基酸的蛋白质，包括一个中断的 1502 个残基的胶原蛋白结构域和一个 232 个残基的 C 端非胶原蛋白结构域。该转录本的表达模式主要在套膜和血细胞中发现。对应于α1(IV)C 端结构域的重组蛋白使成纤维细胞增殖增加了 69%，并使原代培养中产生的胶原蛋白合成增加了一倍。这项工作提供了软体动物非纤维胶原蛋白链的第一个完整的一级结构，以及其 C 端结构域对人细胞的生物学效应。在这项研究中，我们证明了来自软体动物胶原蛋白的 NC1 结构域可以改善人成纤维细胞的增殖和分化。
