title: High resolution spectroscopy reveals fibrillation inhibition pathways of insulin.
abstract: Fibril formation implies the conversion of a protein's native secondary structure and is associated with several neurodegenerative diseases. A better understanding of fibrillation inhibition and fibril dissection requires nanoscale molecular characterization of amyloid structures involved. Tip-enhanced Raman scattering (TERS) has already been used to chemically analyze amyloid fibrils on a sub-protein unit basis. Here, TERS in combination with atomic force microscopy (AFM), and conventional Raman spectroscopy characterizes insulin assemblies generated during inhibition and dissection experiments in the presence of benzonitrile, dimethylsulfoxide, quercetin, and β-carotene. The AFM topography indicates formation of filamentous or bead-like insulin self-assemblies. Information on the secondary structure of bulk samples and of single aggregates is obtained from standard Raman and TERS measurements. In particular the high spatial resolution of TERS reveals the surface conformations associated with the specific agents. The insulin aggregates formed under different inhibition and dissection conditions can show a similar morphology but differ in their β-sheet structure content. This suggests different aggregation pathways where the prevention of the β-sheet stacking of the peptide chains plays a major role. The presented approach is not limited to amyloid-related reasearch but can be readily applied to systems requiring extremely surface-sensitive characterization without the need of labels.
authors.affiliation: Leibniz Institute of Photonic Technology (IPHT), Albert-Einsteinstr. 9, D-07745 Jena, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Deckert-Gaudig
authors.affiliation: Leibniz Institute of Photonic Technology (IPHT), Albert-Einsteinstr. 9, D-07745 Jena, Germany.|Institute of Physical Chemistry, Friedrich-Schiller-University Jena and Abbe Center of Photonics, Helmholtzweg 4, D-07743 Jena, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Deckert
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D009570:Nitriles / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D017433:Protein Structure, Secondary; D011794:Quercetin / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013059:Spectrum Analysis, Raman; D013696:Temperature; D019207:beta Carotene / Q000737:chemistry
chemical_list: D000682:Amyloid; D007328:Insulin; D009570:Nitriles; D010455:Peptides; D012997:Solvents; D019207:beta Carotene; D011794:Quercetin; C014356:benzonitrile; D004121:Dimethyl Sulfoxide