Chaperone-like activity of bovine lens alpha-crystallin in the presence of dithiothreitol-destabilized proteins: characterization of the formed complexes.
Since alpha-crystallin was shown to have in vitro chaperone-like activity, its structure-function relationship has been extensively studied. However, the mechanism of this function is poorly understood. In this study, we monitored the interaction of alpha-crystallin with different proteins namely the insulin B-chain (3.382 kDa), lysozyme (14.4 kDa), and conalbumin (86.18 kDa), all destabilized by dithiothreitol. We found that at 4 degrees C alpha-crystallin prevents the aggregation of destabilized insulin. During the time course of the interaction of alpha-crystallin with the substrate protein, we observed three classes of molecules: the monomeric protein and monomeric alpha-crystallin peptides, alpha-crystallin/substrate protein complexes with a size comparable to alpha-crystallin and large particles. The latter are responsible for the increase of the light scattering of solutions, containing alpha-crystallin/destabilized protein complexes. The molecular exchange between the different populations is temperature dependent and seems to be ruled by the kinetics of the structural changes of the destabilized proteins. At the end all monomeric species are transformed to larger aggregates. The large complexes are enriched in destabilized proteins, compared to the initial ratio alpha-crystallin/substrate protein.
Abgar
S
S
Department of Biochemistry, Biophysics Research Group, University of Antwerp, Universiteitsplein 1, Antwerp, B-2610, Belgium.
Yevlampieva
N
N
Aerts
T
T
Vanhoudt
J
J
Clauwaert
J
J
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D003459:Crystallins / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology*; D007908:Lens, Crystalline / Q000378:metabolism*; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D013054:Spectrophotometry, Atomic; D014461:Ultracentrifugation
D003459:Crystallins; D018832:Molecular Chaperones; D004229:Dithiothreitol