Creating biomimetic surfaces through covalent and oriented binding of proteins

Autor: Jesús M. de la Fuente, Sébastien Chevalier, Mónica Luna, Helene Feracci, Valeria Grazú, Carlos Cuestas-Ayllon
Přispěvatelé: Université Sciences et Technologies - Bordeaux 1, Centre de recherches Paul Pascal (CRPP), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Instituto de Nanociencia de Aragón [Saragoza, España] (INA), University of Zaragoza - Universidad de Zaragoza [Zaragoza], Instituto de Microelectronica de Madrid (IMM), Centro Nacional de Microelectronica [Spain] (CNM)-Consejo Superior de Investigaciones Científicas [Madrid] (CSIC)
Rok vydání: 2010
Předmět:
Nitrilotriacetic Acid
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
02 engineering and technology
law.invention
Mice
law
Biomimetics
Electrochemistry
General Materials Science
Mathematics::Representation Theory
Nuclear Experiment
Cytoskeleton
Spectroscopy
Chelating Agents
0303 health sciences
Chemistry
Surfaces and Interfaces
Adhesion
Hydrogen-Ion Concentration
Silanes
021001 nanoscience & nanotechnology
Condensed Matter Physics
Cadherins
Surface Modification
3. Good health
Membrane
Covalent bond
Metals
0210 nano-technology
Oligopeptides
Protein Binding
Surface Properties
Characterization
Nanotechnology
[SDV.BC]Life Sciences [q-bio]/Cellular Biology
Biofunctionalization
Cell Line
03 medical and health sciences
Confocal microscopy
Cell Adhesion
Animals
Histidine
Cell adhesion
030304 developmental biology
Cadherin
Osmolar Concentration
Immobilized Proteins
Biophysics
Surface modification
Glass
Zdroj: Langmuir
Langmuir, American Chemical Society, 2010, 26 (18), pp.14707-14715. ⟨10.1021/la103086b⟩
Digital.CSIC. Repositorio Institucional del CSIC
instname
ISSN: 1520-5827
0743-7463
DOI: 10.1021/la103086b⟩
Popis: Mónica Luna ...et al.
Mónica Luna...et al.
This manuscript describes a novel method for the biofunctionalization of glass surfaces with polyhistidine-tagged proteins. The main innovation of this methodology consists of the covalent binding between the nitrilotriacetic acid (NTA) moiety and the proteins, ensuring not only orientation, but also stability of the recombinant proteins on NTA-covered surfaces. In this work, as C-terminal polyhistidine tagged cadherin extracellular fragments have been used, this methodology guarantees the proper orientation of these proteins, by mimicking their insertion into cell plasma membranes. These biofunctionalized surfaces have been characterized by confocal microscopy, X-ray photoelectron spectroscopy, contact angle, and atomic force microscopy, showing a high density of cadherins on the glass surfaces and the stability of the linkage. The prepared materials exhibited a high tendency to promote cell spreading, demonstrating the functionality of the protein and the high utility of these biomaterials to promote cell adhesion events. Interestingly, differences in the cytoskeleton organization have been observed in cells adhering to surfaces with no cadherins or with nonoriented cadherins, in comparison to surfaces functionalized with well-oriented cadherins. This method, which allows the robust immobilization of polyhistidine tagged proteins due to their covalent binding and with a defined orientation, may also find particular usefulness in the making of protein biochips, for analysis of protein−protein interactions, as well as structural and single-molecule studies.
This work was supported by institutional funding from CNRS, as well as by grants from Association pour la Recherche sur le Cancer (subvention libre 3135), Région Aquitaine, Fondation pour la Recherche Médicale, la Ligue Contre le Cancer (Dordogne) CNano GSO (HF), and Programme Hubert Curien Picasso (HF/JMF), MITYC (Spain) through the projects CTQ2005-07993-C0202/BQU, CTQ2008-03739/PPQ, NAN2004-09125-C07-02, and PET2007_0315. M. L. acknowledges financial support from CSIC.
Databáze: OpenAIRE
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