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In Vitro Oxidative Crosslinking of Recombinant Barnacle Cyprid Cement Gland Proteins

Cleverley, RM and Webb, DS and Middlemiss, S and Duke, PW and Clare, AS and Okano, K and Harwood, CR and Aldred, N (2021) 'In Vitro Oxidative Crosslinking of Recombinant Barnacle Cyprid Cement Gland Proteins.' Marine Biotechnology, 23 (6). pp. 928-942. ISSN 1436-2228

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Barnacle adhesion is a focus for fouling-control technologies as well as the development of bioinspired adhesives, although the mechanisms remain very poorly understood. The barnacle cypris larva is responsible for surface colonisation. Cyprids release cement from paired glands that contain proteins, carbohydrates and lipids, although further compositional details are scant. Several genes coding for cement gland-specific proteins were identified, but only one of these showed database homology. This was a lysyl oxidase-like protein (lcp_LOX). LOX-like enzymes have been previously identified in the proteome of adult barnacle cement secretory tissue. We attempted to produce recombinant LOX in <jats:italic>E. coli</jats:italic>, in order to identify its role in cyprid cement polymerisation. We also produced two other cement gland proteins (lcp3_36k_3B8 and lcp2_57k_2F5). lcp2_57k_2F5 contained 56 lysine residues and constituted a plausible substrate for LOX. While significant quantities of soluble lcp3_36k_3B8 and lcp2_57k_2F5 were produced in <jats:italic>E. coli</jats:italic>, production of stably soluble lcp_LOX failed. A commercially sourced human LOX catalysed the crosslinking of lcp2_57k_2F5 into putative dimers and trimers, and this reaction was inhibited by lcp3_36k_3B8. Inhibition of the lcp_LOX:lcp2_57k_2F5 reaction by lcp3_36k_3B8 appeared to be substrate specific, with no inhibitory effect on the oxidation of cadaverine by LOX. The results demonstrate a possible curing mechanism for barnacle cyprid cement and, thus, provide a basis for a more complete understanding of larval adhesion for targeted control of marine biofouling and adhesives for niche applications.

Item Type: Article
Uncontrolled Keywords: Barnacle; Cyprid; Adhesion; Cement; Recombinant; Lysyl oxidase
Divisions: Faculty of Science and Health
Faculty of Science and Health > Life Sciences, School of
SWORD Depositor: Elements
Depositing User: Elements
Date Deposited: 11 Nov 2021 15:23
Last Modified: 18 Aug 2022 10:43

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