title: Atomic force microscopy and force spectroscopy on the assessment of protein folding and functionality.
abstract: Atomic force microscopy (AFM) applied to biological systems can, besides generating high-quality and well-resolved images, be employed to study protein folding via AFM-based force spectroscopy. This approach allowed remarkable advances in the measurement of inter- and intramolecular interaction forces with piconewton resolution. The detection of specific interaction forces between molecules based on the AFM sensitivity and the manipulation of individual molecules greatly advanced the understanding of intra-protein and protein-ligand interactions. Apart from the academic interest in the resolution of basic scientific questions, this technique has also key importance on the clarification of several biological questions of immediate biomedical relevance. Force spectroscopy is an especially appropriate technique for "mechanical proteins" that can provide crucial information on single protein molecules and/or domains. Importantly, it also has the potential of combining in a single experiment spatial and kinetic measurements. Here, the main principles of this methodology are described, after which the ability to measure interactions at the single-molecule level is discussed, in the context of relevant protein-folding examples. We intend to demonstrate the potential of AFM-based force spectroscopy in the study of protein folding, especially since this technique is able to circumvent some of the difficulties typically encountered in classical thermal/chemical denaturation studies.
authors.affiliation: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Filomena A
authors.initials: FA
authors.lastname: Carvalho
authors.forename: Ivo C
authors.initials: IC
authors.lastname: Martins
authors.forename: Nuno C
authors.initials: NC
authors.lastname: Santos
mesh_terms: D018625:Microscopy, Atomic Force / Q000379:methods*; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D013057:Spectrum Analysis / Q000379:methods*
chemical_list: D011506:Proteins