Phosphorylation of alpha-crystallin B in Alexander's disease brain.
The phosphorylation of alpha-crystallin B was studied in homogenates of autopsy samples of brain tissue from patients with Alexander's disease, a condition characterized by over-expression of this protein. After incubation in the presence of [gamma-32P]ATP and cAMP the homogenates were analyzed by two-dimensional electrophoresis, (isoelectric focusing followed by SDS-PAGE). Three major polypeptides having the same molecular weight as bovine lens alpha-crystallin B and pIs 7.1, 6.9 and 6.7 were detected in the Coomassie blue stained gels. These three polypeptides were recognized by an alpha-crystallin B-specific antiserum in Western blots. The polypeptides with pIs 7.1 and 6.7 co-migrated in isoelectric focusing gels with bovine lens alpha B and its phosphorylated form alpha Bp, respectively. Radioautography of the two-dimensional gels demonstrated the presence of 32P in the most acidic polypeptide. The results demonstrate the occurrence of alpha B phosphorylation in Alexander's disease brain tissue.
Mann
E
E
Department of Ophthalmology, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
McDermott
M J
MJ
Goldman
J
J
Chiesa
R
R
Spector
A
A
D015153:Blotting, Western; D001923:Brain Chemistry; D003459:Crystallins / Q000032:analysis / Q000378:metabolism*; D002549:Diffuse Cerebral Sclerosis of Schilder / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008970:Molecular Weight; D010750:Phosphoproteins / Q000032:analysis*; D010766:Phosphorylation
D003459:Crystallins; D010750:Phosphoproteins