Phosphorylation of alpha-crystallin in rat lenses is stimulated by H2O2 but phosphorylation has no effect on chaperone activity.
Alpha crystallin (alpha), a phosphorylated structural protein of the lens, has been shown to be a chaperone preventing other lens proteins from aggregating. It is now demonstrated that with oxidative stress imposed on cultured rat lenses, the incorporation of labeled phosphate into the alpha polypeptide chains increased by two to four times over a 90-min period in comparison to control experiments. The phosphorylation rate of the B chain, alpha B, was twice that of the A chain, alpha A. However, phosphorylation of the alpha chains has an insignificant effect on the chaperone activity of alpha or the individual alpha A and alpha B chains as measured by suppressing the thermally induced aggregation of beta low or gamma crystallins. It was also found that the alpha A aggregates are more effective chaperones than the alpha B aggregates. The size of the macromolecules resulting from reaggregation of the isolated non-phosphorylated or phosphorylated alpha B chains are not markedly effected by phosphorylation. However, phosphorylation of the alpha A chain leads to a heterogeneous population with two major species, one similar in size to alpha A and another approximately twice as large. It is concluded that the phosphorylation of alpha is associated with some other function of the protein than that of chaperone activity and that this function may be linked to a protective response to oxidative stress.
Wang
K
K
Department of Ophthalmology, College of Physicians and Surgeons of Columbia University, New York, NY 10032, USA.
Ma
W
W
Spector
A
A
D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007525:Isoelectric Focusing; D007908:Lens, Crystalline / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism*; D018384:Oxidative Stress; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D003459:Crystallins; D018832:Molecular Chaperones; D006861:Hydrogen Peroxide