In situ mapping of biomineral skeletal proteins by molecular recognition imaging withantibody-functionalized AFM tips
In: ISSN: 1742-7061 ; Acta Biomaterialia ; https://hal.science/hal-04272655 ; Acta Biomaterialia, 2023, 168, pp.198-209. ⟨10.1016/j.actbio.2023.07.028⟩ ; https://www.sciencedirect.com/science/article/abs/pii/S1742706123004208, 2023
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Zugriff:
International audience ; Spatial localizing of skeletal proteins in biogenic minerals remains a challenge in biomineralization research. To address this goal, we developed a novel in situ mapping technique based on molecular recognition measurements via atomic force microscopy (AFM), which requires three steps: (1) the development and purification of a polyclonal antibody elicited against the target protein, (2) its covalent coupling to a silicon nitride AFM tip ('functionalization'), and (3) scanning of an appropriately prepared biomineral surface. We applied this approach to a soluble shell protein - accripin11 - recently identified as a major component of the calcitic prisms of the fan mussel Pinna nobilis [1]. Multiple tests reveal that accripin11 is evenly distributed at the surface of the prisms and also present in the organic sheaths surrounding the calcitic prisms, indicating that this protein is both intra- and inter-crystalline. We observed that the adhesion force in transverse sections is about twice higher than in longitudinal sections, suggesting that accripin11 may exhibit preferred orientation in the biomineral. To our knowledge, this is the first time that a protein is localized by molecular recognition atomic force microscopy with antibody-functionalized tips in a biogenic mineral. The 'pros' and 'cons' of this methodology are discussed in comparison with more 'classical' approaches like immunogold. This technique, which leaves the surface to analyze clean, might prove useful for clinical tests on non-pathological (bone, teeth) or pathological (kidney stone) biomineralizations. Studies using implants with protein-doped calcium phosphate coating can also benefit from this technology. STATEMENT OF SIGNIFICANCE: Our paper deals with an unconventional technical approach for localizing proteins that are occluded in biominerals. This technique relies on the use of molecular recognition atomic force microscopy with antibody-functionalized tips. Although such approach has been employed in other system, ...
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In situ mapping of biomineral skeletal proteins by molecular recognition imaging withantibody-functionalized AFM tips
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Autor/in / Beteiligte Person: | Khurshid, Benazir ; Lesniewska, Eric ; Polacchi, Luca ; L'Héronde, Maëva ; Jackson, Daniel J. ; Motreuil, Sébastien ; Thomas, Jérôme ; Bardeau, Jean-François ; Wolf, Stephan E. ; Vielzeuf, Daniel ; Perrin, Jonathan ; Marin, Frédéric ; Biogéosciences UMR 6282 (BGS) ; Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS) ; Synchrotron SOLEIL (SSOLEIL) ; Centre National de la Recherche Scientifique (CNRS) ; Laboratoire Interdisciplinaire Carnot de Bourgogne (ICB) ; Université de Technologie de Belfort-Montbeliard (UTBM)-Université de Bourgogne (UB)-Université Bourgogne Franche-Comté COMUE (UBFC)-Centre National de la Recherche Scientifique (CNRS) ; Institut photonique d'analyse non-destructive européen des matériaux anciens (IPANEMA) ; Muséum national d'Histoire naturelle (MNHN)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Centre National de la Recherche Scientifique (CNRS)-Ministère de la Culture (MC) ; Georg-August-University = Georg-August-Universität Göttingen ; Institut des Molécules et Matériaux du Mans (IMMM) ; Le Mans Université (UM)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS) ; Friedrich-Alexander Universität Erlangen-Nürnberg = University of Erlangen-Nuremberg (FAU) ; Institute of Ceramics and Glass ; Centre Interdisciplinaire de Nanoscience de Marseille (CINaM) ; Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS) ; Two SRO projects, one in 2017, one in 2020 (’Pinocchio’, PInna NObilis & sa Calcification Coquillière: une Histoire Originale) attributed to FM by OSU-Theta (Bourgogne - Franche-Comté), 3) a grant from EMBRC (European Marine Biological Resource Centre, FM), which provided the logistic support for a field mission to collect living specimens of Pinna nobilis at Villefranche-sur-Mer marine station in spring 2017. ; ANR-18-CE02-0014,MOBi,Interface Organique/Inorganique dans les Biominéraux(2018) ; ANR-11-INBS-0004,E-RECOLNAT,Valorisation de 350 ans de collections d'histoire naturelle : une plateforme numérique(2011) |
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Zeitschrift: | ISSN: 1742-7061 ; Acta Biomaterialia ; https://hal.science/hal-04272655 ; Acta Biomaterialia, 2023, 168, pp.198-209. ⟨10.1016/j.actbio.2023.07.028⟩ ; https://www.sciencedirect.com/science/article/abs/pii/S1742706123004208, 2023 |
Veröffentlichung: | HAL CCSD ; Elsevier, 2023 |
Medientyp: | academicJournal |
DOI: | 10.1016/j.actbio.2023.07.028 |
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