Identification and functional characterization of a novel barnacle cement protein

Abstract

Barnacle attachment to various foreign materials in water is guided by an extracellular multiprotein complex. A 19 kDa cement protein was purified from the Megabalanus rosa cement, and its cDNA was cloned and sequenced. The gene was expressed only in the basal portion of the animal, where the histologically identified cement gland is located. The sequence of the protein showed no homology to other known proteins in the databases, indicating that it is a novel protein. Agreement between the molecular mass determined by MS and the molecular weight estimated from the cDNA indicated that the protein bears no post‐translational modifications. The bacterial recombinant was prepared in soluble form under physiologic conditions, and was demonstrated to have underwater irreversible adsorption activity to a variety of surface materials, including positively charged, negatively charged and hydrophobic ones. Thus, the function of the protein was suggested to be coupling to foreign material surfaces during underwater attachment. Homologous genes were isolated from Balanus albicostatus and B. improvisus, and their amino acid compositions showed strong resemblance to that of M. rosa, with six amino acids, Ser, Thr, Ala, Gly, Val and Lys, comprising 66–70% of the total, suggesting that such a biased amino acid composition may be important for the function of this protein.

Micro- and nanostructure of the adhesive material secreted by the tube feet of the sea star Asterias rubens

“…The proteins constituting the adhesive have been characterized in the attachment plaque of mussels and in the cement of barnacles (see Kamino, 2008 andSilverman andRoberto, 2007 for reviews). Those constituting the priming film, rich in polar and charged residues, displace water from the substratum, form strong adhesive bonds with the surface, and provide cohesive coupling with the other components of the adhesive material (Waite and Qin, 2001;Lin et al, 2007;Urushida et al, 2007). Analogous components are probably involved in the formation of the homogeneous film within the adhesive of A. rubens.…”

Temperature Affects Adhesion of the Acorn Barnacle (Balanus amphitrite)

“…The protein contents of barnacle cement has been the topic of considerable research as of late (Kamino 2001;Mori et al 2007;Urushida et al 2007;Kamino 2008;Dickinson et al 2009) yet few cement proteins have been added to the database (19kDa, 20kDa, and 100kDa as well as insoluble partial peptide fractions). Although many were collected from a different, though similar species, Megabalanus rosa, we were consequently hopeful in identifying similar cement polymers on the basis that partial sequence data may be used.…”

“…A particularly interesting study was conducted by Urushida et al (2007) This research has brought to light how a model organism in the laboratory may not translate to a model system for proteomics research. This integrative approach to studying antifouling technologies is imperative to understanding the complete interaction between fouling organism and antifouling or foul-release coating.…”

Barnacle biology before, during and after settlement and metamorphosis: a study of the interface

“…While the patchy material was first observed with the nucleic acid stains, the protein stains suggest the material was mainly proteinaceous and at least some proteins were phosphorylated. Cyprid adhesive and adult glues contain a large fraction of protein (Kamino and Shizuri, 1998;Urushida et al, 2007;He et al, 2013;Gohad et al, 2014). Phosphoprotein was of particular interest as it was found in the cyprid adhesive plaque (Gohad et al, 2014) and is a critical component of other marine adhesives such as mussel (Waite and Qin, 2001;Flammang et al, 2009).…”