The glycation of bovine lens betaL-, betaS- and gamma-crystallins demonstrated by isoelectric focusing and lectin staining.
The aim of the current study is to detect glycation of betaL-, betaS- and gamma-crystallins in the young bovine lens. To determine which of the crystallins are glycated, we have made isoelectric focusing of the water-soluble crystallins of four bovine lenses of 1. 183+/-0.070 years. Samples are stained: (1) with Coomassie Brilliant Blue for proteins; (2) with the lectin Concanavalin-A, followed by horse-radish peroxidase (HRP) and diaminobenzidine (DAB). Experiments are performed with crystallins in native form, in absence of denaturants. The crystallins are separated by isoelectric focusing into: alpha-crystallins of high-molecular weight (HM)-, alphaL-, betaH-, betaL-, betaS- and gamma-crystallins. In the lectin staining experiments only HM-, beta L-, betaS- and gamma-crystallins are positive, whereas the alphaL- and betaH-crystallins do not stain. Though glycation in the bovine lens is very low, lectin staining is sufficiently sensitive to detect the various glycated crystallins.
Ahrend
M H
MH
Institute of Experimental Ophthalmology, University of Bonn, Bonn, D-53105, Germany.
Bours
J
J
D000818:Animals; D002417:Cattle; D003208:Concanavalin A; D003459:Crystallins / Q000032:analysis / Q000378:metabolism*; D006031:Glycosylation; D007525:Isoelectric Focusing; D013194:Staining and Labeling
D003459:Crystallins; D003208:Concanavalin A