title: Effects of monopropanediamino-β-cyclodextrin on the denaturation process of the hybrid protein BlaPChBD.
abstract: Irreversible accumulation of protein aggregates represents an important problem both in vivo and in vitro. The aggregation of proteins is of critical importance in a wide variety of biomedical situations, ranging from diseases (such as Alzheimer's and Parkinson's diseases) to the production (e.g. inclusion bodies), stability, storage and delivery of protein drugs. β-Cyclodextrin (β-CD) is a circular heptasaccharide characterized by a hydrophilic exterior and a hydrophobic interior ring structure. In this research, we studied the effects of a chemically modified β-CD (BCD07056), on the aggregating and refolding properties of BlaPChBD, a hybrid protein obtained by inserting the chitin binding domain of the human macrophage chitotriosidase into the class A β-lactamase BlaP from Bacillus licheniformis 749/I during its thermal denaturation. The results show that BCD07056 strongly increases the refolding yield of BlaPChBD after thermal denaturation and constitutes an excellent additive to stabilize the protein over time at room temperature. Our data suggest that BCD07056 acts early in the denaturation process by preventing the formation of an intermediate which leads to an aggregated state. Finally, the role of β-CD derivatives on the stability of proteins is discussed.
authors.affiliation: Centre d'Ingénierie des Protéines, Université de Liège, Institut de Chimie B6, Sart-Tilman, Liège, Belgium. marylene.vandevenne@sydney.edu.au
authors.forename: Marylène
authors.initials: M
authors.lastname: Vandevenne
authors.forename: Gilles
authors.initials: G
authors.lastname: Gaspard
authors.forename: El Mustapha
authors.initials: el M
authors.lastname: Belgsir
authors.forename: Manilduth
authors.initials: M
authors.lastname: Ramnath
authors.forename: Yves
authors.initials: Y
authors.lastname: Cenatiempo
authors.forename: Daniel
authors.initials: D
authors.lastname: Marechal
authors.forename: Mireille
authors.initials: M
authors.lastname: Dumoulin
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Frere
authors.forename: André
authors.initials: A
authors.lastname: Matagne
authors.forename: Moreno
authors.initials: M
authors.lastname: Galleni
authors.forename: Patrice
authors.initials: P
authors.lastname: Filee
mesh_terms: D002686:Chitin / Q000737:chemistry*; D004795:Enzyme Stability; D011489:Protein Denaturation; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D047392:beta-Cyclodextrins / Q000737:chemistry*; D001618:beta-Lactamases / Q000737:chemistry*
chemical_list: D011993:Recombinant Fusion Proteins; D047392:beta-Cyclodextrins; D002686:Chitin; D001618:beta-Lactamases; C031215:betadex