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Advanced glycation end product ligands for the receptor for advanced glycation end products: Biochemical characterization and formation kinetics

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Author: Valencia, J.V. · Weldon, S.C. · Quinn, D. · Kiers, G.H. · Groot, J. de · TeKoppele, J.M. · Hughes, T.E.
Institution: Gaubius instituut TNO
Source:Analytical Biochemistry, 1, 324, 68-78
Identifier: 237573
doi: doi:10.1016/j.ab.2003.09.013
Keywords: Health Biology · Biomedical Research · Advanced glycation end products · Age · Formation kinetics · Glycation · Maillard reaction · RAGE · Receptor for advanced glycation end products · Serum albumin · 6 n carboxymethyllysine · Advanced glycation end product · Advanced glycation end product receptor · Aldehyde derivative · Bovine serum albumin · Carbonyl derivative · Glyoxylic acid · Ligand · Pentosidine · Ribose · Binding affinity · Chemical reaction kinetics · Correlation analysis · Fluorescence · Glycation · Human cell · Incubation time · Molecular interaction · Molecular mechanics · Nucleotide sequence · Priority journal · Receptor affinity · Receptor binding · Acetaldehyde · Amines · Arginine · Diabetes Mellitus · Fructose · Glucose · Glycosylation End Products, Advanced · Glyoxylates · Humans · Ligands · Lysine · Membrane Proteins · Molecular Weight · Receptors, Immunologic · Recombinant Proteins · Regression Analysis · Ribose · Serum Albumin, Bovine · Spectrometry, Fluorescence · Bovinae


Advanced glycation end products (AGEs) accumulate with age and at an accelerated rate in diabetes. AGEs bind cell-surface receptors including the receptor for advanced glycation end products (RAGE). The dependence of RAGE binding on specific biochemical characteristics of AGEs is currently unknown. Using standardized procedures and a variety of AGE measures, the present study aimed to characterize the AGEs that bind to RAGE and their formation kinetics in vitro. To produce AGEs with varying RAGE binding affinity, bovine serum albumin (BSA) AGEs were prepared with 0.5M glucose, fructose, or ribose at times of incubation from 0 to 12 weeks or for up to 3 days with glycolaldehyde or glyoxylic acid. The AGE-BSAs were characterized for RAGE binding affinity, fluorescence, absorbance, carbonyl content, reactive free amine content, molecular weight, pentosidine content, and N-ε-carboxymethyl lysine content. Ribose-AGEs bound RAGE with high affinity within 1 week of incubation in contrast to glucose- and fructose-AGE, which required 12 and 6 weeks, respectively, to generate equivalent RAGE ligands (IC50=0.66, 0.93, and 1.7μM, respectively). Over time, all of the measured AGE characteristics increased. However, only free amine content robustly correlated with RAGE binding affinity. In addition, detailed protocols for the generation of AGEs that reproducibly bind RAGE with high affinity were developed, which will allow for further study of the RAGE-AGE interaction. © 2003 Elsevier Inc. All rights reserved. Chemicals / CAS: advanced glycation end product receptor, 198785-73-8, 247590-69-8; fructose, 30237-26-4, 57-48-7, 7660-25-5, 77907-44-9; glucose, 50-99-7, 84778-64-3; glyoxylic acid, 298-12-4; pentosidine, 124505-87-9; ribose, 34466-20-1, 50-69-1, 93781-19-2; Acetaldehyde, 75-07-0; advanced glycosylation end-product receptor; Amines; Arginine, 74-79-3; Fructose, 30237-26-4; Glucose, 50-99-7; glycolaldehyde, 141-46-8; Glycosylation End Products, Advanced; Glyoxylates; glyoxylic acid, 298-12-4; Ligands; Lysine, 56-87-1; Membrane Proteins; N(6)-carboxymethyllysine, 5746-04-3; pentosidine, 124505-87-9; Receptors, Immunologic; Recombinant Proteins; Ribose, 50-69-1; Serum Albumin, Bovine Molecular Sequence Numbers: GENBANK: CAA76847;