Bovine Serum Albumin protofibril-like aggregates formation: Solo but not simple mechanism

6533b7dafe1ef96bd126eb26

RESEARCH PRODUCT

Bovine Serum Albumin protofibril-like aggregates formation: Solo but not simple mechanism

Andrea Ponzoni Vito Foderà Giorgio Sberveglieri Valeria Militello Valeria Militello Maurizio Leone Maurizio Leone Valeria Vetri Michele D'amico

subject

Circular dichroism Protein Conformation Globular protein Static Electricity Biophysics Protein aggregation Biochemistry protein aggregation amyloid fibril fluorescence conformational changes chemistry.chemical_compound Protein structure Animals Benzothiazoles Bovine serum albumin Molecular Biology chemistry.chemical_classification biology Temperature Tryptophan Serum Albumin Bovine Hydrogen-Ion Concentration Settore FIS/07 - Fisica Applicata(Beni Culturali Ambientali Biol.e Medicin)Kinetics Thiazoles Crystallography Isoelectric point chemistry Protein destabilization biology.protein Thermodynamics Cattle Thioflavin Protein Multimerization

description

We report an experimental study on the model protein Bovine Serum Albumin (BSA), with the aim of elucidating the mechanisms by which a fully folded globular protein undergoes different aggregation pathways leading to the formation of amyloid fibrils or amorphous aggregates. We observe thermally induced formation of fibrillar structures at pH far from the protein isoelectric point. The increase of electrostatic repulsion results in protein destabilization and in modifications of inter and intra-molecular interactions leading to the growth of fibril-like aggregates stabilized by inter-molecular-β sheets. The aggregation kinetics is studied by means of fluorescence techniques, light scattering, Circular Dichroism (CD), infrared spectroscopy (FTIR) and Atomic Force Microscopy (AFM). Changes in protein secondary structures turn out to be the driving mechanism of the observed aggregation and they progress in parallel with the growth of Thioflavin T emission intensity and scattering signal. This concurrent behavior suggests a mutual stabilization of elongated protofibril-like structures and of protein conformational and structural changes, which lead to a more rigid and ordered structures. Our results give new insights on BSA self-assembly process in alkaline conditions clearly providing new pieces of evidences of the interplay of several and interconnected mechanisms occurring on different time and length scales.

year	journal	country	edition	language
2011-01-01

10.1016/j.abb.2011.01.024 http://hdl.handle.net/11379/157809