Early glycation products produce pentosidine cross-links on native proteins. novel mechanism of pentosidine formation and propagation of glycation.
Bovine lens alpha-crystallin was immobilized on EAH-Sepharose gel and glycated using d-ribose. Incubation with 500 and 100 mm d-ribose for 2 and 15 days produced short-term glycated (STGP gel) and long-term glycated proteins (LTGP gel). Both STGP and LTGP gels produced oxygen free radicals. Hydroxyl radical production was twice that in STGP gel compared with the LTGP gel. Incubation with the glycated gels produced pentosidine in a mixture of N-alpha-acetylarginine + N-alpha-acetyllysine, bovine lens proteins (BLP), and lysozyme; the amounts measured with STGP gel were higher than those with LTGP gel. Reactive oxygen species scavengers decreased the formation of pentosidine. Pentosidine was also formed in BLP when incubated with water-insoluble proteins extracted from aged or brunescent human lenses. Early glycated proteins from aged or diabetic lenses were bound to a boronate affinity column, the protein-containing gel was incubated with BLP, and pentosidine was measured in the incubation mixtures. With this method we found that diabetic lens proteins produced more pentosidine on BLP than did aged lens proteins. Further investigation indicates that two and three carbon carbohydrates possibly formed from oxidative cleavage of early glycation products are involved in pentosidine formation. Based on our findings, we propose a novel pathway for pentosidine formation on native proteins from glycated proteins.
Chellan
P
P
Center for Vision Research, Department of Ophthalmology, Case Western Reserve University and University Hospitals of Cleveland, Cleveland, Ohio 44106, USA.
Nagaraj
R H
RH
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry*; D002241:Carbohydrates / Q000737:chemistry; D002417:Cattle; D002614:Chelating Agents / Q000737:chemistry; D003459:Crystallins / Q000737:chemistry; D005947:Glucose / Q000737:chemistry*; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D017382:Reactive Oxygen Species
D002241:Carbohydrates; D002614:Chelating Agents; D003459:Crystallins; D011506:Proteins; D017382:Reactive Oxygen Species; C002315:N(alpha)-acetyllysine; D001120:Arginine; C062187:pentosidine; D009113:Muramidase; D005947:Glucose; D008239:Lysine; D010100:Oxygen; C000015:N-acetyl-L-arginine