The molecular chaperone, alpha-crystallin, protects against loss of antigenicity and activity of esterase caused by sugars, sugar phosphate and a steroid.
Previously we showed that glycation-induced inactivation and loss of antigenicity of enzymes occur simultaneously. Alpha-crystallin, a major structural protein of the mammalian lens, prevents the aggregation of other proteins and protects enzyme function against post-translational modification in vitro. However, it is not known whether alpha-crystallin can also protect against loss of antigenicity of enzymes. Esterase activity in the lens is decreased in senile cataract and diabetes. We investigated the loss of antigenicity of esterase caused by different insults and the ability of alpha-crystallin to protect. Inactivation of carboxylesterase by sugars, fructose 6-phosphate (F6P) and a steroid, prednisolone-21-hemisuccinate (P-21-H), was measured spectrophotometrically in the presence and absence of alpha-crystallin, while loss of antigenicity was monitored simultaneously using an immunoprecipitation method. The esterase was progressively inactivated by fructose, F6P, ribose, and P-21-H. Bovine alpha-crystallin fully protected against inactivation of esterase by all four compounds, and also protected against loss of antigenicity of the esterase by fructose, ribose and P-21-H at a molar ratio of 1:1. The results indicated that alpha-crystallin, under our experimental conditions, clearly exhibited the ability to prevent loss of antigenicity and inactivation of esterase. The protective effect of alpha-crystallin against loss of antigenicity indicates a novel aspect of its chaperoning function.
Yan
Hong
H
Nuffield Laboratory of Ophthalmology, University of Oxford, Walton St., Oxford OX2 6AW, UK.
Harding
John J
JJ
D000818:Animals; D002241:Carbohydrates / Q000276:immunology*; D043182:Carboxylesterase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000276:immunology / Q000494:pharmacology; D005632:Fructose / Q000494:pharmacology; D005636:Fructosephosphates / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007908:Lens, Crystalline / Q000737:chemistry / Q000378:metabolism; D011233:Precipitin Tests; D011239:Prednisolone / Q000031:analogs & derivatives* / Q000494:pharmacology*; D012266:Ribose / Q000494:pharmacology; D038201:alpha-Crystallins / Q000502:physiology*
D002241:Carbohydrates; D004791:Enzyme Inhibitors; D005636:Fructosephosphates; D038201:alpha-Crystallins; D005632:Fructose; C027618:fructose-6-phosphate; D012266:Ribose; D011239:Prednisolone; D043182:Carboxylesterase; C021322:prednisolone hemisuccinate; D005947:Glucose