In vitro dephosphorylation of alpha-crystallin is dependent on the state of oligomerization

alphaA- and alphaB-crystallin, members of the small heat shock protein fami ly, are present in lens cell extracts as large aggregates. Both alpha -crys tallins are found partially phosphorylated. This study tests the ability of five phosphatases (protein phosphatase PP1, PP2A, PP2B, alkaline and acid phosphatases) to dephosphorylate aA- and alphaB-crystallin in vitro. Activi ty of a phosphatase was dependent on the size of the aggregate. Each of the phosphatases tested showed different specificity and efficiency towards al phaA- and alphaB-crystallins, which depended on the oligomeric state of the alpha -crystallin aggregate. Alkaline phosphatase dephosphorylated both al phaA- and alphaB-crystallin. The reaction was faster when alpha -crystallin was in a tetrameric form. PP2A dephosphorylated primarily alphaA-crystalli n but only after the conversion of alpha -crystallin to tetramers. PPI and PP2B did not dephosphorylate either alphaA- or alphaB-crystallins present a s large aggregates but could not be tested on the lower molecular weight fo rm of alphaA-crystallin. Acid phosphatase dephosphorylated both alphaA- and alphaB-crystallin. The results suggest that an important relationship exis ts between the structure of alpha -crystallin and its level of phosphorylat ion in the cell. (C) 2001 Elsevier Science B.V. All rights reserved.