Prolonged exposure (>90 days) of bovine β-lactoglobulin (BLG) to subdenaturing concentrations of either urea or potassium thiocyanate resulted in the formation of ordered polymers in the form of fibrils. The fibrils obtained with each chaotrope showed major differences in morphology, surface properties, thiol accessibility, and stability to dissociating agents as a consequence of the different chemical bonds involved in their stabilization. Hydrophobic interactions between BLG monomers are predominant in thiocyanate-formed fibrils, whereas urea-formed fibrils are stabilized by intermolecular disulfides generated through a thiol-disulfide exchange reaction. The different features of fibrils obtained with each chaotrope relate to the peculiar structural features and chemical properties of the “active” monomers generated by subdenaturing chaotrope concentrations in the early phases of the polymerization process, as detected by spectroscopic and limited proteolysis/mass spectrometry studies in the earliest stages of the action of individual chaotropes. The chaotrope-specific features of these early intermediates in turn affect the polymerization mechanism, whose intermediates were studied by size-exclusion chromatography on the soluble fraction at different times of fibril formation. The potential of these findings for the production of protein-derived nanostructures having different and controlled geometries and chemical properties is also discussed.
|Titolo:||Formation of structured polymers upon controlled denaturation of beta-lactoglobulin with different chaotropes|
|Autori interni:||BONOMI, FRANCESCO|
BARBIROLI, ALBERTO GIUSEPPE (Secondo)
RASMUSSEN, PATRIZIA STELLA SKOV (Primo)
IAMETTI, STEFANIA (Ultimo)
|Parole Chiave:||bovine β-lactoglobulin ; polymers ; fibrils ; urea ; thiocyanate|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore AGR/12 - Patologia Vegetale
|Data di pubblicazione:||2007|
|Digital Object Identifier (DOI):||10.1002/bip.20704|
|Appare nelle tipologie:||01 - Articolo su periodico|
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