Temperature dependence of chaperone-like activity and oligomeric state of alphaB-crystallin.
The chaperone-like activity and the oligomeric state of alphaB-crystallin were studied at different temperatures and in the presence of urea and thiocyanate. The activity, assessed measuring the ability of alphaB-crystallin to prevent the aggregation of denatured insulin, strongly depends on temperature. While a significant activity increase was detected at 42 degrees C, the presence of urea and thiocyanate does not affect the protein activity in an irreversible way. In-solution SAXS measurements performed in the same experimental conditions showed that alphaB-crystallin forms near-spherical, hollowed, polydisperse oligomers, whose dimensions change above 42 degrees C. Moreover, in the presence of urea and thiocyanate, a global fit analysis confirms the high stability of alphaB-crystallin assemblies in relationship with their variable quaternary structure. In particular, the changes in the inner radius as well as the thickness and dispersion of the protein shell, account for the preservation of the chaperone-like activity.
Spinozzi
Francesco
F
Dipartimento di Scienze applicate ai Sistemi Complessi, Università Politecnica delle Marche, and INFM Unità di Ancona, Via Brecce Bianche, I-60131 Ancona, Italy. f.spinozzi@alisf1.univpm.it
Mariani
Paolo
P
Rustichelli
Franco
F
Amenitsch
Heinz
H
Bennardini
Federico
F
Mura
Giovanni Maria
GM
Coi
Alessio
A
Ganadu
Maria Luisa
ML
D019281:Dimerization; D006360:Heat-Shock Proteins / Q000737:chemistry; D006801:Humans; D007328:Insulin / Q000737:chemistry; D018832:Molecular Chaperones / Q000502:physiology*; D010940:Plant Proteins / Q000737:chemistry; D020836:Protein Structure, Quaternary; D013696:Temperature; D013861:Thiocyanates / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology; D038203:alpha-Crystallin B Chain / Q000737:chemistry* / Q000502:physiology*
C101303:HSP16.9 protein, Triticum aestivum; D006360:Heat-Shock Proteins; D007328:Insulin; D018832:Molecular Chaperones; D010940:Plant Proteins; D013861:Thiocyanates; D038203:alpha-Crystallin B Chain; D014508:Urea; C031760:thiocyanate