Mucins and molluscan calcification -: Molecular characterization of mucoperlin, a novel mucin-like protein from the nacreous shell layer of the fan mussel Pinna nobilis (Bivalvia, Pteriomorphia)
Mucins and molluscan calcification -: Molecular characterization of mucoperlin, a novel mucin-like protein from the nacreous shell layer of the fan mussel Pinna nobilis (Bivalvia, Pteriomorphia)
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DOI：
10.1074/jbc.m003006200
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发表时间：
2000-07-07
影响因子：
4.8
通讯作者：
Westbroek, P
中科院分区：
文献类型：
--
作者：
Marin, F;Corstjens, P;Westbroek, P
一个cDNA表达文库构建的外套膜组织的mRNA的地中海扇贻贝Pinna nobilis进行了筛选，提出了对乙酸可溶性壳基质的同一物种的抗体。这导致了一个2138碱基对的cDNA的分离，含有93个碱基对的13个串联重复。推导的蛋白质具有66.7 kDa的分子量和4.8的等电点。这种蛋白质，这是富含丝氨酸和脯氨酸残基，过表达，纯化，并用于生产多克隆抗体。原位和体外免疫测试表明，该蛋白质定位于P. nobilis的珍珠层文石层中，但不在钙化棱镜中。由于P. nobilis珍珠层的这种蛋白质表现出一些粘蛋白样特征，我们提出了名称mucoperlin，这是第一篇报道软体动物粘蛋白克隆的论文，也是第一个粘蛋白参与软体动物钙化的分子证据。这一发现证实了我们先前的假设，即软体动物壳基质的一些蛋白质成分将来自粘蛋白，这是晚前寒武纪的许多后生动物谱系所共有的（Marin，F.，史密斯，M.，伊萨，Y.，Muyzer，G.和Westbroek，P.（1996）Proc. Natl，Acad，Sci，U. S. A. 93，1554-1559），适应一个祖先粘蛋白的新功能，矿化过程的调节，可能是其中一个分子事件，这将解释在寒武纪大爆发期间，在许多后生动物谱系中同时出现有组织的钙化。
A cDNA expression library constructed from mantle tissue mRNA of the Mediterranean fan mussel Pinna nobilis was screened with antibodies raised against the acetic acid-soluble shell matrix of the same species. This resulted in the isolation of a 2138-base pair cDNA, containing 13 tandem repeats of 93 base pairs. The deduced protein has a molecular mass of 66.7 kDa and a isoelectric point of 4.8. This protein, which is enriched in serine and proline residues, was overexpressed, purified, and used for producing polyclonal antibodies. Immunological in situ and in vitro tests showed that the protein is localized in the nacreous aragonitic layer of P. nobilis, but not in the calcitic prisms, Because this protein of the nacre of P nobilis exhibits some mucin-like characteristics, we propose the name mucoperlin, This is the first paper reporting the cloning of a molluscan mucin and the first molecular evidence for the involvement of a mucin in molluscan calcification. This finding corroborates our previous hypothesis that some of the proteinaceous constituents of the molluscan shell matrix would derive from mucins, common to many metazoan lineages of the late Precambrian (Marin, F., Smith, M., Isa, Y., Muyzer, G. and Westbroek, P. (1996) Proc. Natl, Acad, Sci, U. S. A. 93, 1554-1559), The adaptation of an ancestral mucin to a new function, the regulation of the mineralization process, may be one of the molecular events, among others, that would explain the simultaneous emergence of organized calcification in many metazoan lineages during the Cambrian explosion.