title: Advanced glycation end product ligands for the receptor for advanced glycation end products: biochemical characterization and formation kinetics.
abstract: 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-epsilon-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 1.7 microM, 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.
authors.affiliation: Novartis Institutes for BioMedical Research, 100 Technology Square, Bldg. 601/Rm. 5155, Cambridge, MA 02139, USA.
authors.forename: Jessica V
authors.initials: JV
authors.lastname: Valencia
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Weldon
authors.forename: Douglas
authors.initials: D
authors.lastname: Quinn
authors.forename: Geesje H
authors.initials: GH
authors.lastname: Kiers
authors.forename: Jeroen
authors.initials: J
authors.lastname: DeGroot
authors.forename: Johan M
authors.initials: JM
authors.lastname: TeKoppele
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Hughes
mesh_terms: D000079:Acetaldehyde / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D000588:Amines / Q000032:analysis; D001120:Arginine / Q000031:analogs & derivatives* / Q000032:analysis; D003920:Diabetes Mellitus / Q000378:metabolism; D005632:Fructose / Q000737:chemistry / Q000378:metabolism; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000138:chemical synthesis / Q000378:metabolism*; D006038:Glyoxylates / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012044:Regression Analysis; D012266:Ribose / Q000737:chemistry / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D000588:Amines; D017127:Glycation End Products, Advanced; D006038:Glyoxylates; D008024:Ligands; D008565:Membrane Proteins; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D005632:Fructose; D012266:Ribose; C048496:N(6)-carboxymethyllysine; D001120:Arginine; C062187:pentosidine; D000079:Acetaldehyde; D005947:Glucose; C031150:glyoxylic acid; D008239:Lysine; C010972:glycolaldehyde