title: Surface attachment of protein fibrils via covalent modification strategies.
abstract: Chemical control of surface functionality and topography is an essential requirement for many technological purposes. In particular, the covalent attachment of monomeric proteins to surfaces has been the object of intense studies in recent years, for applications as varied as electrochemistry, immuno-sensing, and the production of biocompatible coatings. Little is known, however, about the characteristics and requirements underlying surface attachment of supramolecular protein nanostructures. Amyloid fibrils formed by the self-assembly of peptide and protein molecules represent one important class of such structures. These highly organized beta-sheet-rich assemblies are a hallmark of a range of neurodegenerative disorders, including Alzheimer's disease and type II diabetes, but recent findings suggest that they have much broader significance, potentially representing the global free energy minima of the energy landscapes of proteins and having potential applications in material science. In this paper, we describe strategies for attaching amyloid fibrils formed from different proteins to gold surfaces under different solution conditions. Our methods involve the reaction of sulfur containing small molecules (cystamine and 2-iminothiolane) with the amyloid fibrils, enabling their covalent linkage to gold surfaces. We demonstrate that irreversible attachment using these approaches makes possible quantitative analysis of experiments using biosensor techniques, such as quartz crystal microbalance (QCM) assays that are revolutionizing our understanding of the mechanisms of amyloid growth and the factors that determine its kinetic behavior. Moreover, our results shed light on the nature and relative importance of covalent versus noncovalent forces acting on protein superstructures at metal surfaces.
authors.affiliation: Nanoscience Centre, University of Cambridge, 11 JJ Thomson Avenue, CB3 0FF, UK.
authors.forename: Alexander K
authors.initials: AK
authors.lastname: Buell
authors.forename: Duncan A
authors.initials: DA
authors.lastname: White
authors.forename: Christoph
authors.initials: C
authors.lastname: Meier
authors.forename: Mark E
authors.initials: ME
authors.lastname: Welland
authors.forename: Tuomas P J
authors.initials: TP
authors.lastname: Knowles
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D015374:Biosensing Techniques; D003538:Cystamine / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D018625:Microscopy, Atomic Force; D011506:Proteins / Q000737:chemistry
chemical_list: D000682:Amyloid; D011506:Proteins; D006046:Gold; D003538:Cystamine