Characterization of Two 20kDa-Cement Protein (cp20k) Homologues in Amphibalanus amphitrite
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Abstract
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. © 2013 He et al.Citation
He L-S, Zhang G, Qian P-Y (2013) Characterization of Two 20kDa-Cement Protein (cp20k) Homologues in Amphibalanus amphitrite. PLoS ONE 8: e64130. doi:10.1371/journal.pone.0064130.Sponsors
The authors’ research grant from China Ocean Mineral Resources Research and Development Association (DY125-15-T-02), and King Abdullah University of Science and Technology (SA-C0040/UK-C0016) to P.Y. Qian. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Publisher
Public Library of Science (PLoS)Journal
PLoS ONEISSN
1932-6203PubMed ID
23717550PubMed Central ID
PMC3661472Handle URI
http://hdl.handle.net/10754/334590Scopus Count
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