title: Experimental strategies for the identification and characterization of adhesive proteins in animals: a review.
abstract: Adhesive secretions occur in both aquatic and terrestrial animals, in which they perform diverse functions. Biological adhesives can therefore be remarkably complex and involve a large range of components with different functions and interactions. However, being mainly protein based, biological adhesives can be characterized by classical molecular methods. This review compiles experimental strategies that were successfully used to identify, characterize and obtain the full-length sequence of adhesive proteins from nine biological models: echinoderms, barnacles, tubeworms, mussels, sticklebacks, slugs, velvet worms, spiders and ticks. A brief description and practical examples are given for a variety of tools used to study adhesive molecules at different levels from genes to secreted proteins. In most studies, proteins, extracted from secreted materials or from adhesive organs, are analysed for the presence of post-translational modifications and submitted to peptide sequencing. The peptide sequences are then used directly for a BLAST search in genomic or transcriptomic databases, or to design degenerate primers to perform RT-PCR, both allowing the recovery of the sequence of the cDNA coding for the investigated protein. These sequences can then be used for functional validation and recombinant production. In recent years, the dual proteomic and transcriptomic approach has emerged as the best way leading to the identification of novel adhesive proteins and retrieval of their complete sequences.
authors: Biology of Marine Organisms and Biomimetics, Research Institute for Biosciences , University of Mons , 23 Place du Parc, 7000 Mons , Belgium.
authors: Elise
authors: E
authors: Hennebert
authors: Molecular Biology and Genetic Engineering, GIGA-R , University of Liège , 1 Avenue de l'Hôpital, 4000 Liège , Belgium.
authors: Barbara
authors: B
authors: Maldonado
authors: Institute of Zoology and Center of Molecular Bioscience Innsbruck , University of Innsbruck , Technikerstrasse 25, 6020 Innsbruck , Austria.
authors: Peter
authors: P
authors: Ladurner
authors: Biology of Marine Organisms and Biomimetics, Research Institute for Biosciences , University of Mons , 23 Place du Parc, 7000 Mons , Belgium.
authors: Patrick
authors: P
authors: Flammang
authors: Unidade de Investigação em Ciências Orais e Biomédicas, Faculdade de Medicina Dentária , Universidade de Lisboa, Cidade Universitária , 1649-003 Lisboa , Portugal.
authors: Romana
authors: R
authors: Santos
keywords: biological adhesion; metazoans; protein characterization
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