title: β-Connectin studies by small-angle x-ray scattering and single-molecule force spectroscopy by atomic force microscopy.
abstract: The three-dimensional structure and the mechanical properties of a β-connectin fragment from human cardiac muscle, belonging to the I band, from I(27) to I(34), were investigated by small-angle x-ray scattering (SAXS) and single-molecule force spectroscopy (SMFS). This molecule presents an entropic elasticity behavior, associated to globular domain unfolding, that has been widely studied in the last 10 years. In addition, atomic force microscopy based SMFS experiments suggest that this molecule has an additional elastic regime, for low forces, probably associated to tertiary structure remodeling. From a structural point of view, this behavior is a mark of the fact that the eight domains in the I(27)-I(34) fragment are not independent and they organize in solution, assuming a well-defined three-dimensional structure. This hypothesis has been confirmed by SAXS scattering, both on a diluted and a concentrated sample. Two different models were used to fit the SAXS curves: one assuming a globular shape and one corresponding to an elongated conformation, both coupled with a Coulomb repulsion potential to take into account the protein-protein interaction. Due to the predominance of the structure factor, the effective shape of the protein in solution could not be clearly disclosed. By performing SMFS by atomic force microscopy, mechanical unfolding properties were investigated. Typical sawtooth profiles were obtained and the rupture force of each unfolding domain was estimated. By fitting a wormlike chain model to each peak of the sawtooth profile, the entropic elasticity of octamer was described.
authors: Department of Physics, University of Florence and CNISM, Sesto Fiorentino (Florence), Italy.
authors: S
authors: S
authors: Marchetti
authors: F
authors: F
authors: Sbrana
authors: A
authors: A
authors: Toscano
authors: E
authors: E
authors: Fratini
authors: M
authors: M
authors: Carlà
authors: M
authors: M
authors: Vassalli
authors: B
authors: B
authors: Tiribilli
authors: A
authors: A
authors: Pacini
authors: C M C
authors: CM
authors: Gambi
mesh_terms: D064211:Connectin; D004548:Elasticity; D006801:Humans; D018625:Microscopy, Atomic Force; D009124:Muscle Proteins / Q000737:chemistry*; D011494:Protein Kinases / Q000737:chemistry*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D058767:Protein Unfolding; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
chemical_list: D064211:Connectin; D009124:Muscle Proteins; C577285:TTN protein, human; D011494:Protein Kinases