A novel chitin-binding protein from the vestimentiferan Riftia pachyptila interacts specifically with β-chitin -: Cloning, expression, and characterization
A novel chitin-binding protein from the vestimentiferan Riftia pachyptila interacts specifically with β-chitin -: Cloning, expression, and characterization
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DOI：
10.1074/jbc.m009244200
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发表时间：
2001-03-16
影响因子：
4.8
通讯作者：
Delachambre, J
中科院分区：
文献类型：
--
作者：
Chamoy, L;Nicolaï, M;Delachambre, J
克隆了一个来自厚膜里夫蒂亚菌（Riftia pachyptila）的cDNA。它编码一种新的21.3 kDa的蛋白质，从蠕虫的保护管，命名为RCBP（为里夫蒂亚几丁质结合蛋白）。在先前获得的部分管肽序列的基础上，使用逆转录酶介导的聚合酶链反应和cDNA末端的快速扩增的实验导致了完整的cDNA序列。其推导的氨基酸序列的分析表明存在两个几丁质结合结构域。这些结构域与仅限于动物界的2型几丁质结合结构域密切相关。我们通过亲和测定和免疫金标记表明RCBP是迄今已知的第一种特异性结合β-几丁质的蛋白质，并且不能结合在几丁质分泌动物中发现的最常见的α-形式。RCBP mRNA被发现存在于特定的表皮细胞从蠕虫体壁，但从来没有在几丁质分泌腺细胞。这个意想不到的结果清楚地表明，这种管蛋白是在外上皮的专门区域合成的，并且至少有两种不同的组织参与这种外骨骼合成。
A cDNA from Riftia pachyptila was cloned. It encodes a novel 21.3-kDa protein from the worm protective tube, named RCBP (for Riftia chitin-binding protein). On the basis of partial tube-peptide sequences previously obtained, experiments using reverse transcriptase-mediated polymerase chain reaction and rapid amplification of cDNA ends led to the complete cDNA sequence. Analysis of its deduced amino acid sequence shows the presence of two chitin-binding domains. These domains are closely related to type 2 chitin-binding domains that are restricted to the animal kingdom. We showed by affinity assay and immunogold labeling that RCBP is the first protein so far known that binds specifically beta -chitin and that is unable to bind the most common alpha -form found in chitin secreting animals. The RCBP mRNA was found to be present in specific epidermal cells from the worm body wall, but never in the chitin-secreting gland cells. This unexpected result clearly indicates that this tube protein is synthesized in specialized areas of the outer epithelium and that at least two different tissues are involved in this exoskeleton synthesis.