Inactivation and loss of antigenicity of esterase by sugars and a steroid.
Glycation, the non-enzymic reaction of sugars with proteins, has an important role in the complications of diabetes. It has been studied mostly in structural proteins but more recently has been shown to inactivate enzymes. Previous evidence from our laboratory indicated that glycation-induced inactivation and loss of antigenicity of catalase and superoxide dismutase are simultaneous. Esterase, which decreases activity in the lens in senile cataract and diabetes, was measured by a spectrophotometric assay using p-nitrophenyl acetate as the substrate. Here we investigated the inactivation of carboxylesterase (EC 3.1.1.1) by sugars of different glycating power and prednisolone-21-hemisuccinate while simultaneously monitoring the loss of antigenicity. Antigenicity was assessed by immunoprecipitation and by dot-blotting the glycated and non-glycated fractions of enzymes separated by affinity chromatography. Ribose and fructose inactivated more rapidly than glucose and glucose 6-phosphate. The esterase was progressively inactivated by prednisolone-21-hemisuccinate at a lower concentration. Activity and antigenicity were lost simultaneously. The glycated enzyme had entirely lost its antigenicity. These results further support the idea that inactivation of enzyme and loss of antigenicity are simultaneous.
Yan
H
H
Nuffield Laboratory of Ophthalmology, University of Oxford, Walton St., Oxford OX2 6AW, UK.
Harding
J J
JJ
D000375:Aging / Q000378:metabolism; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity; D003920:Diabetes Mellitus / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects*; D005632:Fructose / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D019298:Glucose-6-Phosphate / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D011233:Precipitin Tests; D011239:Prednisolone / Q000031:analogs & derivatives / Q000494:pharmacology; D012266:Ribose / Q000494:pharmacology
D005632:Fructose; D019298:Glucose-6-Phosphate; D012266:Ribose; D011239:Prednisolone; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; C021322:prednisolone hemisuccinate; D005947:Glucose