Localization of Phosphoproteins within the Barnacle Adhesive Interface.
Localization of Phosphoproteins within the Barnacle Adhesive Interface.
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DOI：
10.1086/bblv230n3p233
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发表时间：
2016-06
期刊：
影响因子：
--
通讯作者：
Beniash E
中科院分区：
文献类型：
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作者：
Dickinson GH;Yang X;Wu F;Orihuela B;Rittschof D;Beniash E
藤壶使用蛋白质胶永久粘附在几乎任何惰性基质上。这种胶由至少十种主要蛋白质组成，其中一些已被分离和测序。关于粘合所涉及的化学机制以及胶水作为钙化基板矿化平台的潜力仍然存在疑问。我们测试了藤壶胶含有磷蛋白的假设，磷蛋白有可能在粘附和矿化中发挥作用。通过结合磷蛋白特异性凝胶染色和抗磷酸丝氨酸抗体的蛋白质印迹，我们在藤壶 Amphibalanus amphitrite 的未固化胶分泌物中鉴定出多种磷酸化蛋白。不同藤壶的胶蛋白组成以及磷蛋白的数量和丰度存在明显差异，这可能是由于胶液分泌随时间的周期性变化所致。我们使用脱钙的藤壶基板和重新附着的藤壶沉积的残留胶来评估磷蛋白在藤壶胶层内的位置。在基板的有机基质和残留的胶水中发现了磷蛋白。残留胶内的染色在毛细胶管（参与胶的循环释放）已铺设的区域中出现得最强烈。最后，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离的胶蛋白的矿化研究表明，被鉴定为磷酸化的蛋白质可能诱导碳酸钙（CaCO3）的矿化。这些结果有助于我们了解藤壶胶的蛋白质组成，并为磷蛋白在水下生物粘合剂中的潜在作用提供新的见解。
Barnacles permanently adhere to nearly any inert substrate using proteinaceous glue. The glue consists of at least ten major proteins, some of which have been isolated and sequenced. Questions still remain about the chemical mechanisms involved in adhesion and the potential of the glue to serve as a platform for mineralization of the calcified base plate. We tested the hypothesis that barnacle glue contains phosphoproteins, which have the potential to play a role in both adhesion and mineralization. Using a combination of phosphoprotein-specific gel staining and Western blotting with anti-phosphoserine antibody, we identified multiple phosphorylated proteins in uncured glue secretions from the barnacle Amphibalanus amphitrite. The protein composition of the glue and the quantity and abundance of phosphoproteins varied distinctly among individual barnacles, possibly due to cyclical changes in the glue secretion over time. We assessed the location of the phosphoproteins within the barnacle glue layer using decalcified barnacle base plates and residual glue deposited by reattached barnacles. Phosphoproteins were found throughout the organic matrix of the base plate and within the residual glue. Staining within the residual glue appeared most intensely in regions where capillary glue ducts, which are involved in cyclical release of glue, had been laid down. Lastly, mineralization studies of glue proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that proteins identified as phosphorylated possibly induce mineralization of calcium carbonate (CaCO3). These results contribute to our understanding of the protein composition of barnacle glue, and provide new insights into the potential roles of phosphoproteins in underwater bioadhesives.