title: Amyloid fibril formation by bovine α1-acid glycoprotein in a reducing environment: The role of disulfide bridges on the observed aggregation kinetics.
abstract: Bovine α1-acid glycoprotein (bAGP), a thermostable counterpart of its human homologue, is a positive acute phase protein involved in binding and transportation of a large number of bio-active molecules and drugs across the body. We have investigated the effect of low pH and reducing conditions on the structure of the protein and found that it aggregates at high temperatures. The aggregates show a fibrillar structure when observed with electron microscopy. Aggregation assays using the amyloid-specific dye Thioflavin T show the presence of a lag phase which was neither abolished nor shortened when seeds were added. A priori reduction of the two disulfide bridges of bAGP, on the other hand, abolished the lag phase and reveals a connection between the kinetics of reduction and aggregation. We provide a kinetic interpretation and the corresponding rate laws allowing to model the process of fibril formation by bAGP under reducing conditions. Our interpretation allows to assess the role of disulfide bridges on the fibrillation kinetics of bAGP and can provide a more accurate interpretation of the fibrillation kinetics of other amyloidogenic proteins containing disulfide bridges.
authors.affiliation: Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden. Electronic address: maurizio.baldassarre@dbb.su.se.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Baldassarre
authors.affiliation: Department of Life and Environmental Sciences, Università Politecnica delle Marche, Ancona, Italy.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Maggiore
authors.affiliation: Department of Life and Environmental Sciences, Università Politecnica delle Marche, Ancona, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Scirè
authors.affiliation: Department of Life and Environmental Sciences, Università Politecnica delle Marche, Ancona, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Tanfani
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002417:Cattle; D004220:Disulfides / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D009961:Orosomucoid / Q000378:metabolism*; D010084:Oxidation-Reduction; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D000682:Amyloid; D004220:Disulfides; D009961:Orosomucoid
keywords: Amyloid-like fibrils; Disulfide bridge; Infrared spectroscopy; Orosomucoid; Protein aggregation; α(1)-acid glycoprotein