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Characterization of Two 20kDa-Cement Protein (cp20k) Homologues in Amphibalanus amphitrite

机译：角闪石两性的两个20kDa水泥蛋白（cp20k）同源物的表征

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摘要

The barnacle, Amphibalanus amphitrite, is a common marine fouling organism. Understanding the mechanism of barnacle adhesion will be helpful in resolving the fouling problem. Barnacle cement is thought to play a key role in barnacle attachment. Although several adult barnacle cement proteins have been identified in Megabalanus rosa, little is known about their function in barnacle settlement. In this study, two homologous 20k-cement proteins (cp20k) in Amphibalanus amphitrite, named Bamcp20k-1 and Bamcp20k-2, were characterized. The two homologues share primary sequence structure with proteins from other species including Megabalanus rosa and Fistulobalanus albicostatus. The conserved structure included repeated Cys domains and abundant charged amino acids, such as histidine. In this study we demonstrated that Bamcp20k-1 localized at the α secretory cells in the cyprid cement gland, while Bamcp20k-2 localized to the β secretory cells. The differential localizations suggest differential regulation for secretion from the secretory cells. Both Bamcp20k-1 and Bamcp20k-2 from cyprids dissolved in PBS. However, adult Bamcp20k-2, which was dominant in the basal shell of adult barnacles, was largely insoluble in PBS. Solubility increased in the presence of the reducing reagent Dithiothreitol (DTT), suggesting that the formation of disulfide bonds plays a role in Bamcp20k-2 function. In comparison, Bamcp20k-1, which was enriched in soft tissue, could not be easily detected in the shell and base by Western blot and easily dissolved in PBS. These differential solubilities and localizations indicate that Bamcp20k-1 and Bamcp20k-2 have distinct functions in barnacle cementing.
机译：藤壶Amphibalanus amphitrite是一种常见的海洋污染生物。了解藤壶粘附的机制将有助于解决结垢问题。藤壶水泥被认为在藤壶附着中起关键作用。尽管已经在罗氏沼虾中鉴定了几种成年的藤壶粘固蛋白，但对其在藤壶沉降中的功能了解甚少。在这项研究中，表征了在两栖闪石两栖动物中的两个同源20k水泥蛋白（cp20k），分别称为Bamcp20k-1和Bamcp20k-2。这两个同源物与来自其他物种的蛋白质（包括罗汉果和拟南芥）共享蛋白质的一级序列结构。保守的结构包括重复的Cys结构域和大量带电荷的氨基酸，例如组氨酸。在这项研究中，我们证明了Bamcp20k-1定位在塞浦路斯水泥腺的α分泌细胞中，而Bamcp20k-2定位于β分泌细胞中。不同的定位提示从分泌细胞分泌的不同调节。塞浦路斯的Bamcp20k-1和Bamcp20k-2均溶于PBS。然而，在成年藤壶的基壳中占优势的成年Bamcp20k-2基本上不溶于PBS。在还原剂二硫苏糖醇（DTT）的存在下，溶解度增加，这表明二硫键的形成在Bamcp20k-2功能中起作用。相比之下，富含软组织的Bamcp20k-1难以通过Western blot在壳和碱基中检测出来，并且很容易溶于PBS。这些不同的溶解度和位置表明Bamcp20k-1和Bamcp20k-2在藤壶固井中具有不同的功能。

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    Li-Sheng He; Gen Zhang; Pei-Yuan Qian;

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  • 年(卷),期 -1(8),5
  • 年度 -1
  • 页码 e64130
  • 总页数 9
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