title: Oxidase Activity of the Barnacle Adhesive Interface Involves Peroxide-Dependent Catechol Oxidase and Lysyl Oxidase Enzymes.
abstract: Oxidases are found to play a growing role in providing functional chemistry to marine adhesives for the permanent attachment of macrofouling organisms. Here, we demonstrate active peroxidase and lysyl oxidase enzymes in the adhesive layer of adult Amphibalanus amphitrite barnacles through live staining, proteomic analysis, and competitive enzyme assays on isolated cement. A novel full-length peroxinectin (AaPxt-1) secreted by barnacles is largely responsible for oxidizing phenolic chemistries; AaPxt-1 is driven by native hydrogen peroxide in the adhesive and oxidizes phenolic substrates typically preferred by phenoloxidases (POX) such as laccase and tyrosinase. A major cement protein component AaCP43 is found to contain ketone/aldehyde modifications via 2,4-dinitrophenylhydrazine (DNPH) derivatization, also called Brady's reagent, of cement proteins and immunoblotting with an anti-DNPH antibody. Our work outlines the landscape of molt-related oxidative pathways exposed to barnacle cement proteins, where ketone- and aldehyde-forming oxidases use peroxide intermediates to modify major cement components such as AaCP43.
authors.forename: Christopher R
authors.identifier: http://orcid.org/0000-0001-7572-778X
authors.initials: CR
authors.lastname: So
authors.forename: Jenifer M
authors.initials: JM
authors.lastname: Scancella
authors.forename: Kenan P
authors.initials: KP
authors.lastname: Fears
authors.forename: Tara
authors.initials: T
authors.lastname: Essock-Burns
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Haynes
authors.forename: Dagmar H
authors.initials: DH
authors.lastname: Leary
authors.forename: Zoie
authors.initials: Z
authors.lastname: Diana
authors.forename: Chenyue
authors.initials: C
authors.lastname: Wang
authors.forename: Stella
authors.initials: S
authors.lastname: North
authors.forename: Christina S
authors.initials: CS
authors.lastname: Oh
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Nicholas School of the Environment and Earth Sciences, Duke University Marine Laboratory , Beaufort, North Carolina 28516, United States.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Orihuela
authors.affiliation: Nicholas School of the Environment and Earth Sciences, Duke University Marine Laboratory , Beaufort, North Carolina 28516, United States.
authors.forename: Dan
authors.initials: D
authors.lastname: Rittschof
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Spillmann
authors.forename: Kathryn J
authors.identifier: http://orcid.org/0000-0001-8163-6964
authors.initials: KJ
authors.lastname: Wahl
mesh_terms: D000269:Adhesives; D000818:Animals; D004156:Catechol Oxidase; D010088:Oxidoreductases / Q000378:metabolism*; D010545:Peroxides; D008249:Protein-Lysine 6-Oxidase; D040901:Proteomics; D001468:Thoracica
chemical_list: D000269:Adhesives; D010545:Peroxides; D010088:Oxidoreductases; D004156:Catechol Oxidase; D008249:Protein-Lysine 6-Oxidase
keywords: barnacles; bioadhesion; biofouling; catechol oxidation; chemical modification; lysyl oxidase; peroxidase; protein cross-linking