title: Strategies for a direct characterization of phosphoproteins on hydroxyapatite surfaces.
abstract: We show in this work how systems formed by phosphoproteins on calcium phosphate surfaces can be directly characterized, in real time, in liquid medium, without the need for elution or labeling. Specifically, we show how this is possible by applying three different techniques: ellipsometry, quartz crystal microbalance with dissipation, and atomic force microscopy-based friction force spectroscopy. We apply these techniques to study two different model systems, i.e. those formed upon the adsorption of two model phosphoproteins (κ- and β-casein) on hydroxyapatite (HA) surfaces. Information on the kinetics of adsorption, surface excess, viscoelasticity, water content, thickness of the layers, and protein-surface interaction is provided. Results indicate that both phosphoproteins form homogeneous elastic highly hydrated monolayers on the HA surfaces, the strength of β-casein layers being higher by approximately a factor of 4. Based on the experimental results, models for the conformation of κ- and β-casein molecules adsorbed on HA surfaces are proposed.
authors.affiliation: Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö, Sweden.
authors.forename: J
authors.initials: J
authors.lastname: Sotres
authors.forename: A
authors.initials: A
authors.lastname: Barrantes
authors.forename: L
authors.initials: L
authors.lastname: Lindh
authors.forename: T
authors.initials: T
authors.lastname: Arnebrant
mesh_terms: D000327:Adsorption; D002021:Buffers; D002130:Calcium Phosphates / Q000737:chemistry; D002364:Caseins / Q000737:chemistry*; D004058:Diffusion; D017886:Durapatite / Q000737:chemistry*; D055119:Elastic Modulus; D004548:Elasticity; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force; D011487:Protein Conformation; D058525:Quartz Crystal Microbalance Techniques; D013057:Spectrum Analysis / Q000379:methods; D013499:Surface Properties; D013997:Time Factors; D014783:Viscosity; D014867:Water / Q000737:chemistry
chemical_list: D002021:Buffers; D002130:Calcium Phosphates; D002364:Caseins; D014867:Water; D017886:Durapatite; C020243:calcium phosphate