Microbiology, School of Natural Sciences, School of Natural Sciences, National University of Ireland Galway, Galway, Ireland.
Centre for Research in Medical Devices (CÚRAM), School of Natural Sciences, National University of Ireland Galway, Galway, Ireland.
Philos Trans R Soc Lond B Biol Sci. 2019 Oct 28;374(1784):20190205. doi: 10.1098/rstb.2019.0205. Epub 2019 Sep 9.
Adhesive proteins of barnacle cement have potential as environmentally friendly adhesives owing to their ability to adhere to various substrates in aqueous environments. By understanding the taxonomic breath of barnacles with different lifestyles, we may uncover commonalities in adhesives produced by these specialized organisms. The 19 kDa cement protein (cp19k) of the stalked barnacle Pollicipes pollicipes was expressed in Escherichia coli BL21 to investigate its adhesive properties. Initial expression of hexahistidine-tagged protein (rPpolcp19k-his) yielded low levels of insoluble protein. Co-overproduction of E. coli molecular chaperones GroEL-GroES and trigger factor (TF) increased soluble protein yields, although TF co-purified with the target protein (TF-rPpolcp19k-his). Surface coat analysis revealed high levels of adsorption of the TF-rPpolcp19k-his complex and of purified E. coli TF on both hydrophobic and hydrophilic surfaces, while low levels of adsorption were observed for rPpolcp19k-his. Tag-free rPpolcp19k protein also exhibited low adsorption compared to fibrinogen and Cell-Tak controls on hydrophobic, neutral hydrophilic and charged self-assembled monolayers under surface plasmon resonance assay conditions designed to mimic the barnacle cement gland or seawater. Because rPpolcp19k protein displays low adhesive capability, this protein is suggested to confer the ability to self-assemble into a plaque within the barnacle cement complex. This article is part of the theme issue 'Transdisciplinary approaches to the study of adhesion and adhesives in biological systems'.
藤壶黏合蛋白由于其在水相环境中能黏附于多种基质的能力，有望成为环境友好型黏合剂。通过了解不同生活方式的藤壶的分类范围，我们可能会发现这些特殊生物产生的黏合剂具有共性。本文是主题为“生物系统中黏附和黏合剂的跨学科研究方法”的一部分内容。本文研究了具有柄部的藤壶 Pollicipes pollicipes 的 19 kDa 黏合蛋白（cp19k），以调查其黏附特性。最初表达带有六组氨酸标签的蛋白（rPpolcp19k-his）时，得到的不溶性蛋白产量较低。共表达大肠杆菌分子伴侣 GroEL-GroES 和触发因子（TF）增加了可溶性蛋白的产量，尽管 TF 与目标蛋白（TF-rPpolcp19k-his）共纯化。表面涂层分析表明，TF-rPpolcp19k-his 复合物和纯化的大肠杆菌 TF 对疏水性和亲水性表面具有高吸附水平，而 rPpolcp19k-his 的吸附水平较低。无标签的 rPpolcp19k 蛋白的吸附水平也低于纤维蛋白原和 Cell-Tak 对照物，在设计用于模拟藤壶黏合腺或海水的表面等离子体共振分析条件下，疏水、中性亲水和带电荷的自组装单层上的吸附水平较低。由于 rPpolcp19k 蛋白显示出较低的黏附能力，因此该蛋白可能赋予其在藤壶黏合复合物内自我组装成斑块的能力。
