Phosphorylation of alphaB-crystallin alters chaperone function through loss of dimeric substructure.
Phosphorylation is the most common posttranslational modification of the alpha-crystallins in the human lens. These phosphorylated forms are not only important because of their abundance in aging lenses and the implications for cataract but also because they have been identified in patients with degenerative brain disease. By using mimics corresponding to the reported in vivo phosphorylation sites in the human lens, we have examined the effects of phosphorylation upon the chaperone-like properties and structure of alphaB-crystallin. Here we show that phosphorylation of alphaB-crystallin at Ser-45 results in uncontrolled aggregation. By using an innovative tandem mass spectrometry approach, we demonstrate how this alteration in behavior stems from disruption of dimeric substructure within the polydisperse alphaB-crystallin assembly. This structural perturbation appears to disturb the housekeeping role of alphaB-crystallin and consequently has important implications for the disease states caused by protein aggregation in the lens and deposition in non-lenticular tissue.
Aquilina
J Andrew
JA
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
Benesch
Justin L P
JL
Ding
Lin Lin
LL
Yaron
Orna
O
Horwitz
Joseph
J
Robinson
Carol V
CV
D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002942:Circular Dichroism; D019281:Dimerization; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism; D008027:Light; D013058:Mass Spectrometry; D008954:Models, Biological; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000378:metabolism; D009154:Mutation; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D012542:Scattering, Radiation; D012694:Serine / Q000737:chemistry; D013997:Time Factors; D014908:Triticum / Q000378:metabolism; D014466:Ultraviolet Rays; D038203:alpha-Crystallin B Chain / Q000378:metabolism*
D018832:Molecular Chaperones; D038203:alpha-Crystallin B Chain; D012694:Serine