On the composition and origin of the urea-soluble polypeptides of the U18666A cataract.
The composition and origin of the urea soluble polypeptides which accumulate in the U18666A rat-cataract were studied. Chromatography on Sephacryl S-200 in 7.2 M urea separated the USP into 19-20 and 22-26 kDa enriched fractions. The polypeptide composition of these fractions was probed by immunoblotting of IEF and 2-D electrophoresis gels. The cataract USP largely focused at pHs comparable to alpha- and beta-crystallins. Immunoblotting of 2-D gels showed the USP to be composed predominantly of alpha- and beta-derived crystallins; little gamma-polypeptide was detected in the gels. Some of the insoluble alpha-crystallin appeared to be degraded. Changes in the lens WSP which accompanied the increase in USP were also measured. WSP decreased more than USP increased. Decreases in soluble high molecular weight proteins (alpha- plus beta-crystallins) and medium molecular weight proteins (beta-crystallins) were calculated which together could entirely account for the increased USP. An unexpected decrease in the lens soluble low molecular weight proteins (gamma-crystallins) appeared largely due to the selective leakage of gammas from the lens. The protein content of the ocular humors from eyes with cataracts increased 4 fold and contained polypeptides that focused on IEF like gamma-light crystallin and reacted with the gamma-crystallin antiserum. The cause of the protein insolubilization in the U18666A cataract is unknown but could be partially due to increased aggregation of alpha-crystallins secondary to loss of gamma-crystallins from the lens.
Cenedella
R J
RJ
Department of Biochemistry, Kirksville College of Osteopathic Medicine, MO 63501.
Augusteyn
R C
RC
D000736:Androstenes; D000818:Animals; D001082:Aqueous Humor / Q000378:metabolism; D002386:Cataract / Q000139:chemically induced / Q000378:metabolism*; D002850:Chromatography, Gel; D003459:Crystallins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015151:Immunoblotting; D007908:Lens, Crystalline / Q000378:metabolism*; D008297:Male; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014508:Urea
D000736:Androstenes; D003459:Crystallins; D010455:Peptides; D013438:Sulfhydryl Compounds; C006261:3-beta-(2-(diethylamino)ethoxy)androst-5-en-17-one; D014508:Urea