The expression and characterization of recombinant cp19k barnacle cement protein from Pollicipes pollicipes. (28th October 2019)

• Record Type: Journal Article
• Authors: Tilbury, Maura A.; McCarthy, Sean; Domagalska, Magdalena; Ederth, Thomas; Power, Anne Marie; Wall, J. Gerard
• Is Part Of: Philosophical transactions. Volume 374:Number 1784(2019)
• Publication Date: 2019-10-28
• Subjects: barnacle -- adhesion -- protein -- recombinant cp19k -- surface plasmon resonance -- self-assembled monolayers; Biology -- Periodicals; Science -- Periodicals; 570
• DOI: 10.1098/rstb.2019.0205
• Languages: English
• ISSNs: 0962-8436

Abstract

Abstract : 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'.