{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T18:40:43Z","timestamp":1772563243944,"version":"3.50.1"},"reference-count":49,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2021,3,25]],"date-time":"2021-03-25T00:00:00Z","timestamp":1616630400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/BTA-GES\/32359\/2017"],"award-info":[{"award-number":["PTDC\/BTA-GES\/32359\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"European Marine Biology Resource Infrastructure Cluster","award":["ID: 654008"],"award-info":[{"award-number":["ID: 654008"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>We focus on the stalked goose barnacle L. anatifera adhesive system, an opportunistic less selective species for the substrate, found attached to a variety of floating objects at seas. Adhesion is an adaptative character in barnacles, ensuring adequate positioning in the habitat for feeding and reproduction. The protein composition of the cement multicomplex and adhesive gland was quantitatively studied using shotgun proteomic analysis. Overall, 11,795 peptide sequences were identified in the gland and 2206 in the cement, clustered in 1689 and 217 proteinGroups, respectively. Cement specific adhesive proteins (CPs), proteases, protease inhibitors, cuticular and structural proteins, chemical cues, and many unannotated proteins were found, among others. In the cement, CPs were the most abundant (80.5%), being the bulk proteins CP100k and -52k the most expressed of all, and CP43k-like the most expressed interfacial protein. Unannotated proteins comprised 4.7% of the cement proteome, ranking several of them among the most highly expressed. Eight of these proteins showed similar physicochemical properties and amino acid composition to known CPs and classified through Principal Components Analysis (PCA) as new CPs. The importance of PCA on the identification of unannotated non-conserved adhesive proteins, whose selective pressure is on their relative amino acid abundance, was demonstrated.<\/jats:p>","DOI":"10.3390\/ijms22073370","type":"journal-article","created":{"date-parts":[[2021,3,25]],"date-time":"2021-03-25T14:51:12Z","timestamp":1616683872000},"page":"3370","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Proteogenomic Characterization of the Cement and Adhesive Gland of the Pelagic Gooseneck Barnacle Lepas anatifera"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5211-972X","authenticated-orcid":false,"given":"Dany","family":"Dom\u00ednguez-P\u00e9rez","sequence":"first","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9874-933X","authenticated-orcid":false,"given":"Daniela","family":"Almeida","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0320-5883","authenticated-orcid":false,"given":"Josef","family":"Wissing","sequence":"additional","affiliation":[{"name":"Cellular Proteomics Research, Helmholtz Centre for Infection Research, Inhoffenstra\u00dfe. 7, 38124 Braunschweig, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6857-7581","authenticated-orcid":false,"given":"Andr\u00e9 M.","family":"Machado","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"}]},{"given":"Lothar","family":"J\u00e4nsch","sequence":"additional","affiliation":[{"name":"Cellular Proteomics Research, Helmholtz Centre for Infection Research, Inhoffenstra\u00dfe. 7, 38124 Braunschweig, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1328-1732","authenticated-orcid":false,"given":"Agostinho","family":"Antunes","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"},{"name":"Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, s\/n, 4169-007 Porto, Portugal"}]},{"given":"Lu\u00eds Filipe","family":"Castro","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"},{"name":"Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, s\/n, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3585-2417","authenticated-orcid":false,"given":"Vitor","family":"Vasconcelos","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"},{"name":"Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, s\/n, 4169-007 Porto, Portugal"}]},{"given":"Alexandre","family":"Campos","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6149-9840","authenticated-orcid":false,"given":"Isabel","family":"Cunha","sequence":"additional","affiliation":[{"name":"CIIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua General Norton de Matos s\/n, Terminal de Cruzeiros do Porto de Leix\u00f5es, 4450-208 Matosinhos, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,25]]},"reference":[{"key":"ref_1","first-page":"1","article-title":"The cement apparatus of the stalked barnacle Pollicipes pollicipes","volume":"164","author":"Alves","year":"2017","journal-title":"Mar. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1377","DOI":"10.1002\/jmor.20067","article-title":"Unusual adhesive production system in the barnacle Lepas anatifera: An ultrastructural and histochemical investigation","volume":"273","author":"Jonker","year":"2012","journal-title":"J. Morphol."},{"key":"ref_3","unstructured":"Southward, A.J., and Crisp, D.J. (1987). Barnacle ecology and adaptation. Barnacle Biology, A.A. Balkema."},{"key":"ref_4","first-page":"226","article-title":"Relationships between wave exposure and biomass of the goose barnacle (Pollicipes pollicipes, Gmelin, 1790) in the Gaztelugatxe Marine Reserve (Basque Country, northern Spain)","volume":"63","author":"Borja","year":"2016","journal-title":"ICES J. Mar. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1007\/s00253-010-2913-8","article-title":"Protein-based underwater adhesives and the prospects for their biotechnological production","volume":"89","author":"Stewart","year":"2011","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1080\/08927014.2010.542809","article-title":"Economic impact of biofouling on a naval surface ship","volume":"27","author":"Schultz","year":"2011","journal-title":"Biofouling"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/s10126-007-9076-3","article-title":"Underwater Adhesive of Marine Organisms as the Vital Link Between Biological Science and Material Science","volume":"10","author":"Kamino","year":"2008","journal-title":"Mar. Biotechnol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3009","DOI":"10.1021\/acs.biomac.7b00969","article-title":"Degradable Adhesives for Surgery and Tissue Engineering","volume":"18","author":"Bhagat","year":"2017","journal-title":"Biomacromolecules"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3259","DOI":"10.1002\/adfm.201303536","article-title":"Enhancing the Adhesion of a Biomimetic Polymer Yields Performance Rivaling Commercial Glues","volume":"24","author":"Meredith","year":"2014","journal-title":"Adv. Funct. Mater."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Kamino, K. (2006). Barnacle underwater attachment. Biological Adhesives, Springer Berlin Heidelberg.","DOI":"10.1007\/978-3-540-31049-5_8"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Kamino, K. (2016). Barnacle underwater attachment. Biological Adhesives, Springer.","DOI":"10.1007\/978-3-319-46082-6_7"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"36219","DOI":"10.1038\/srep36219","article-title":"Sequence basis of Barnacle Cement Nanostructure is Defined by Proteins with Silk Homology","volume":"6","author":"So","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_13","first-page":"171","article-title":"Transcriptome and Proteome Studies Reveal Candidate Attachment Genes during the Development of the Barnacle Amphibalanus Amphitrite","volume":"3","author":"Ryu","year":"2016","journal-title":"Front. Mar. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Chen, Z.-F., Zhang, H., Wang, H., Matsumura, K., Wong, Y.H., Ravasi, T., and Qian, P.-Y. (2014). Quantitative Proteomics Study of Larval Settlement in the Barnacle Balanus amphitrite. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0088744"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Dominguez-Perez, D., Almeida, D., Wissing, J., Machado, A.M., J\u00e4nsch, L., Castro, L.F., Antunes, A., Vasconcelos, V., Campos, A., and Cunha, I. (2020). The Quantitative Proteome of the Cement and Adhesive Gland of the Pedunculate Barnacle, Pollicipes pollicipes. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072524"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"565","DOI":"10.3389\/fmars.2019.00565","article-title":"Biochemistry of Barnacle Adhesion: An Updated Review","volume":"6","author":"Liang","year":"2019","journal-title":"Front. Mar. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1007\/s10126-018-9856-y","article-title":"Comparative Analysis of the Adhesive Proteins of the Adult Stalked Goose Barnacle Pollicipes pollicipes (Cirripedia: Pedunculata)","volume":"21","author":"Rocha","year":"2019","journal-title":"Mar. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"11493","DOI":"10.1021\/acsami.7b01185","article-title":"Oxidase Activity of the Barnacle Adhesive Interface Involves Peroxide-Dependent Catechol Oxidase and Lysyl Oxidase Enzymes","volume":"9","author":"So","year":"2017","journal-title":"Acs Appl. Mater. Interfaces"},{"key":"ref_19","first-page":"1","article-title":"Acorn Barnacles Secrete Phase-Separating Fluid to Clear Surfaces Ahead of Cement Deposition","volume":"5","author":"Fears","year":"2018","journal-title":"Adv. Sci."},{"key":"ref_20","first-page":"280","article-title":"Cirripedia\u2014non-parasitic Thoracica","volume":"Volume 50","author":"Costello","year":"2001","journal-title":"European Register of Marine Species: A Check-List of the Marine Species in Europe and a Bibliography of Guides to Their Identification. Collection Patrimoines Naturels"},{"key":"ref_21","unstructured":"(2021, February 03). Worms Lepas Anatifera Linnaeus. Available online: https:\/\/www.marinespecies.org\/aphia.php?p=taxdetails&id=106149."},{"key":"ref_22","first-page":"S48","article-title":"Feeding and Spawning of the Goose Barnacle Lepas anatifera (Cirripedia, Lepadidae) on Floating Substrates in the Open Northwestern Pacific Ocean","volume":"81","author":"Reznichenko","year":"2001","journal-title":"Entomol. Rev. C\/C Entomol. Obozr."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.seares.2013.04.005","article-title":"Patterns of distribution and abundance of the stalked barnacle (Pollicipes pollicipes) in the central and southwest coast of continental Portugal","volume":"83","author":"Sousa","year":"2013","journal-title":"J. Sea Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1163\/001121611X574290","article-title":"South African pelagic goose barnacles (Cirripedia, Thoracica): Substratum preferences and influence of plastic debris on abundance and distribution","volume":"84","author":"Whitehead","year":"2011","journal-title":"Crustaceana"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"170","DOI":"10.2307\/1539940","article-title":"Comparative Histological Studies of the Cement Apparatus of Lepas anatifera and Balanus tintinnabulum","volume":"137","author":"Lacombe","year":"1969","journal-title":"Biol. Bull."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/ncomms5414","article-title":"Synergistic roles for lipids and proteins in the permanent adhesive of barnacle larvae","volume":"5","author":"Gohad","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1111\/j.1742-4658.2012.08552.x","article-title":"Significance of the conformation of building blocks in curing of barnacle underwater adhesive","volume":"279","author":"Kamino","year":"2012","journal-title":"FEBS J."},{"key":"ref_28","unstructured":"Vayalam, J.P.V. (2012). Barnacles and Their Significance in Biofouling. Operational and Environmental Consequences of Large Industrial Cooling Water Systems, Springer."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/srep44980","article-title":"First evidence for temporary and permanent adhesive systems in the stalked barnacle cyprid, Octolasmis angulata","volume":"7","author":"Yap","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1017\/S0025315499001617","article-title":"Active relocation in lepadomorph barnacles","volume":"80","author":"Kugele","year":"2000","journal-title":"J. Mar. Biol. Assoc."},{"key":"ref_31","first-page":"1","article-title":"Directional locomotion in a turtle barnacle, Chelonibia testudinaria, on green turtles, Chelonia mydas","volume":"119","author":"Moriarty","year":"2008","journal-title":"Mar. Turt. Newsl."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Ferrier, G.A., Kim, S.J., Kaddis, C.S., Loo, J.A., Ann Zimmer, C., and Zimmer, R.K. (2016, January 3\u20137). Multifuncin: A Multifunctional Protein Cue Induces Habitat Selection by, and Predation on, Barnacles. Proceedings of the Integrative and Comparative Biology, Portland, Oregon.","DOI":"10.1093\/icb\/icw076"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1016\/S0965-1748(00)00062-X","article-title":"Identification of methyl farnesoate in the cypris larva of the barnacle, Balanus amphitrite, and its role as a juvenile hormone","volume":"30","author":"Smith","year":"2000","journal-title":"Insect Biochem. Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1007\/s002270050053","article-title":"Characterization of barnacle (Balanus eburneus and B. cenatus) adhesive proteins","volume":"127","author":"Naldrett","year":"1997","journal-title":"Mar. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"6436","DOI":"10.1111\/j.1742-4658.2007.06161.x","article-title":"Calcite-specific coupling protein in barnacle underwater cement","volume":"274","author":"Mori","year":"2007","journal-title":"FEBS J."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"He, L.-S., Zhang, G., and Qian, P.-Y. (2013). Characterization of Two 20kDa-Cement Protein (cp20k) Homologues in Amphibalanus amphitrite. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0064130"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1080\/08927014.2013.853051","article-title":"First study on gene expression of cement proteins and potential adhesion-related genes of a membranous-based barnacle as revealed from Next-Generation Sequencing technology","volume":"30","author":"Lin","year":"2014","journal-title":"Biofouling"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1093\/protein\/4.2.155","article-title":"Correlation between stability of a protein and its dipeptide composition: A novel approach for predicting in vivo stability of a protein from its primary sequence","volume":"4","author":"Guruprasad","year":"1990","journal-title":"Protein Eng."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/0022-2836(82)90515-0","article-title":"A simple method for displaying the hydropathic character of a protein","volume":"157","author":"Kyte","year":"1982","journal-title":"J. Mol. Biol."},{"key":"ref_40","first-page":"1895","article-title":"Thermostability and aliphatic index of globular proteins","volume":"88","author":"Ikai","year":"1980","journal-title":"J. Biochem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1002\/prot.20587","article-title":"Protein flexibility and rigidity predicted from sequence","volume":"61","author":"Schlessinger","year":"2005","journal-title":"Proteins Struct. Funct. Bioinforma."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.jprot.2016.01.010","article-title":"Shotgun proteomics to unravel marine mussel (Mytilus edulis) response to long-term exposure to low salinity and propranolol in a Baltic Sea microcosm","volume":"137","author":"Campos","year":"2016","journal-title":"J. Proteom."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.ab.2015.08.018","article-title":"Minimizing technical variation during sample preparation prior to label-free quantitative mass spectrometry","volume":"490","author":"Scheerlinck","year":"2015","journal-title":"Anal. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"757","DOI":"10.15252\/msb.20145625","article-title":"Ultrasensitive proteome analysis using paramagnetic bead technology","volume":"10","author":"Hughes","year":"2014","journal-title":"Mol. Syst. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4060","DOI":"10.1021\/acs.jproteome.7b00433","article-title":"Evaluation of FASP, SP3, and iST Protocols for Proteomic Sample Preparation in the Low Microgram Range","volume":"16","author":"Sielaff","year":"2017","journal-title":"J. Proteome Res."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Machado, A.M., Sarropoulou, E., Castro, L.F.C., Vasconcelos, V., and Cunha, I. (2019). An important resource for understanding bio-adhesion mechanisms: Cement gland transcriptomes of two goose barnacles, Pollicipes pollicipes and Lepas anatifera (Cirripedia, Thoracica). Mar. Genom., 45.","DOI":"10.1016\/j.margen.2018.11.001"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1093\/nar\/gku366","article-title":"PredictProtein-an open resource for online prediction of protein structural and functional features","volume":"42","author":"Yachdav","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1098\/rstb.2019.0195","article-title":"Interspecies comparison of sea star adhesive proteins","volume":"374","author":"Lengerer","year":"2019","journal-title":"Philos. Trans. R. Soc. B Biol. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Davey, P.A., Power, A.M., Santos, R., Bertemes, P., Ladurner, P., Palmowski, P., Clarke, J., Flammang, P., Lengerer, B., and Hennebert, E. (2021). Omics-based molecular analyses of adhesion by aquatic invertebrates. Biol. Rev.","DOI":"10.1111\/brv.12691"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/7\/3370\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:41:03Z","timestamp":1760161263000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/7\/3370"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,25]]},"references-count":49,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["ijms22073370"],"URL":"https:\/\/doi.org\/10.3390\/ijms22073370","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,25]]}}}