title: Well-defined biomimetic surfaces to characterize glycosaminoglycan-mediated interactions on the molecular, supramolecular and cellular levels.
abstract: Glycosaminoglycans (GAGs) are ubiquitously present at the cell surface and in extracellular matrix, and crucial for matrix assembly, cell-cell and cell-matrix interactions. The supramolecular presentation of GAG chains, along with other matrix components, is likely to be functionally important but remains challenging to control and to characterize, both in vivo and in vitro. We present a method to create well-defined biomimetic surfaces that display GAGs, either alone or together with other cell ligands, in a background that suppresses non-specific binding. Through the design of the immobilization platform - a streptavidin monolayer serves as a molecular breadboard for the attachment of various biotinylated ligands - and a set of surface-sensitive in situ analysis techniques (including quartz crystal microbalance and spectroscopic ellipsometry), the biomimetic surfaces are tailor made with tight control on biomolecular orientation, surface density and lateral mobility. Analysing the interactions between a selected GAG (heparan sulphate, HS) and the HS-binding chemokine CXCL12α (also called SDF-1α), we demonstrate that these surfaces are versatile for biomolecular and cellular interaction studies. T-lymphocytes are found to adhere specifically to surfaces presenting CXCL12α, both when reversibly bound through HS and when irreversibly immobilized on the inert surface, even in the absence of any bona fide cell adhesion ligand. Moreover, surfaces which present both HS-bound CXCL12α and the intercellular adhesion molecule 1 (ICAM-1) synergistically promote cell adhesion. Our surface biofunctionalization strategy should be broadly applicable for functional studies that require a well-defined supramolecular presentation of GAGs along with other matrix or cell-surface components.
authors.affiliation: Univ. Grenoble Alpes, Departement de Chimie Moléculaire (DCM), Grenoble, France; CNRS, DCM, Grenoble, France.
authors.forename: Elisa
authors.initials: E
authors.lastname: Migliorini
authors.affiliation: Univ. Grenoble Alpes, Departement de Chimie Moléculaire (DCM), Grenoble, France; CNRS, DCM, Grenoble, France.
authors.forename: Dhruv
authors.initials: D
authors.lastname: Thakar
authors.affiliation: Univ. Grenoble Alpes, Institut de Biologie Structurale (IBS), Grenoble, France; CNRS, IBS, Grenoble, France; CEA, DSV, IBS, Grenoble, France.
authors.forename: Rabia
authors.initials: R
authors.lastname: Sadir
authors.affiliation: Department of Cellular Logistics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Tino
authors.initials: T
authors.lastname: Pleiner
authors.affiliation: Unité de Chimie des Biomolécules, Département de Biologie Structurale et Chimie, Institut Pasteur, Paris, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Baleux
authors.affiliation: Univ. Grenoble Alpes, Institut de Biologie Structurale (IBS), Grenoble, France; CNRS, IBS, Grenoble, France; CEA, DSV, IBS, Grenoble, France.
authors.forename: Hugues
authors.initials: H
authors.lastname: Lortat-Jacob
authors.affiliation: Univ. Grenoble Alpes, Departement de Chimie Moléculaire (DCM), Grenoble, France; CNRS, DCM, Grenoble, France.
authors.forename: Liliane
authors.initials: L
authors.lastname: Coche-Guerente
authors.affiliation: Univ. Grenoble Alpes, Departement de Chimie Moléculaire (DCM), Grenoble, France; CNRS, DCM, Grenoble, France; CIC biomaGUNE, Biosurfaces Unit, San Sebastian, Spain; Max Planck Institute for Intelligent Systems, Stuttgart, Germany. Electronic address: rrichter@cicbiomagune.es.
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Richter
mesh_terms: D032701:Biomimetics / Q000379:methods*; D019921:Biotinylation; D002448:Cell Adhesion; D002462:Cell Membrane / Q000737:chemistry*; D054377:Chemokine CXCL12 / Q000737:chemistry*; D005109:Extracellular Matrix / Q000737:chemistry; D005353:Fibronectins / Q000737:chemistry; D006025:Glycosaminoglycans / Q000737:chemistry*; D006497:Heparan Sulfate / Q000737:chemistry; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000737:chemistry*; D019169:Jurkat Cells; D008024:Ligands; D008958:Models, Molecular; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013601:T-Lymphocytes / Q000737:chemistry
chemical_list: D054377:Chemokine CXCL12; D005353:Fibronectins; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D018799:Intercellular Adhesion Molecule-1; D008024:Ligands; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D019809:Streptavidin
keywords: Chemokine; Glycosaminoglycan; Quartz crystal microbalance; Spectroscopic ellipsometry; Surface plasmon resonance; T-lymphocyte adhesion