{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T02:22:38Z","timestamp":1790734958277,"version":"4.1.0"},"reference-count":52,"publisher":"Portland Press Ltd.","issue":"13","license":[{"start":{"date-parts":[[2021,6,11]],"date-time":"2021-06-11T00:00:00Z","timestamp":1623369600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,7,16]]},"abstract":"<jats:p>Centrins are conserved calcium (Ca2+)-binding proteins typically associated with centrosomes that have been implicated in several biological processes. In Toxoplasma gondii, a parasite that causes toxoplasmosis, three centrin isoforms have been recognized. We have recently characterized the metal binding and structural features of isoform 1 (TgCEN1), demonstrating that it possesses properties consistent with a role as a Ca2+ sensor and displays a Ca2+-dependent tendency to self-assemble. Herein, we expanded our studies, focusing on the self-association and target binding properties of TgCEN1 by combining biophysical techniques including dynamic light scattering, isothermal titration calorimetry, nuclear magnetic resonance, circular dichroism, and fluorescence spectroscopy. We found that the self-assembly process of TgCEN1 depends on different physicochemical factors, including Ca2+ concentration, temperature, and protein concentration, and is mediated by both electrostatic and hydrophobic interactions. The process is completely abolished upon removal of the first 21-residues of the protein and is significantly reduced in the presence of a binding target peptide derived from the human XPC protein (P17-XPC). Titration of P17-XPC to the intact protein and isolated domains showed that TgCEN1 possesses two binding sites with distinct affinities and Ca2+ sensitivity; a high-affinity site in the C-lobe which may be constitutively bound to the peptide and a low-affinity site in the N-lobe which is active only upon Ca2+ stimulus. Overall, our results suggest a specific mechanism of TgCEN1 for Ca2+-modulated target binding and support a N-to-C self-assembly mode, in which the first 21-residues of one molecule likely interact with the C-lobe of the other.<\/jats:p>","DOI":"10.1042\/bcj20210295","type":"journal-article","created":{"date-parts":[[2021,6,11]],"date-time":"2021-06-11T09:37:23Z","timestamp":1623404243000},"page":"2571-2587","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["The interplay of self-assembly and target binding in centrin 1 from <i>Toxoplasma gondii<\/i>"],"prefix":"10.1042","volume":"478","author":[{"given":"Carolina","family":"Conter","sequence":"first","affiliation":[{"name":"Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Bombardi","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Pedretti","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Filippo","family":"Favretto","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adele","family":"Di Matteo","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology, CNR, Piazzale Aldo Moro, 5, 00185 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Dominici","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7341-0970","authenticated-orcid":false,"given":"Alessandra","family":"Astegno","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2021,7,9]]},"reference":[{"key":"2021112218224479300_BCJ-2021-0295C1","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1111\/j.1550-7408.1992.tb01468.x","article-title":"Characterization of the calcium-binding contractile protein centrin from Tetraselmis striata (Pleurastrophyceae)","volume":"39","year":"1992","journal-title":"J. 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