title: Structural and immunological characterization of Amadori-rich human serum albumin: role in diabetes mellitus.
abstract: Proteins modifications in diabetes may lead to early glycation products (EGPs) as well as advanced glycation end products (AGEs). Whereas no extensive studies have been carried out to assess the role of EGPs in secondary complications of diabetes, numerous investigators have demonstrated the role of AGEs. Early glycation involves attachment of glucose on ε-NH2 of lysine residues of proteins leading to generation of the Amadori product (an early glycation species). This study reports the structural and immunological characterization of EGPs of HSA because we believe that during persistent hyperglycemia the HSA, one of the major blood proteins, can undergo fast glycation. Glucose mediated generation of EGPs of HSA was quantitated as Amadori products by NBT assay and authenticated by boronate affinity chromatography and LC/MS. Compared to native HSA changes in glycated-HSA were characterized by hyperchromicity, loss in fluorescence intensity and a new peak in the FTIR profile. Immunogenicity of native- and glycated-HSA was evaluated by inducing antibodies in rabbits. Results suggest generation of neo-epitopes on glycated-HSA rendering it highly immunogenic compared to native HSA. Quantization of EGPs of HSA by authentic antibodies against HSA-EGPs can be used as marker for early detection of the initiation/progression of secondary complications of diabetes.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, A.M.U., Aligarh 202002, UP, India.
authors.forename: Binish
authors.initials: B
authors.lastname: Arif
authors.forename: Jalaluddin M
authors.initials: JM
authors.lastname: Ashraf
authors.lastname: Moinuddin
authors.forename: Jamal
authors.initials: J
authors.lastname: Ahmad
authors.forename: Zarina
authors.initials: Z
authors.lastname: Arif
authors.forename: Khursheed
authors.initials: K
authors.lastname: Alam
mesh_terms: D002846:Chromatography, Affinity; D003922:Diabetes Mellitus, Type 1 / Q000097:blood*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D017127:Glycation End Products, Advanced / Q000096:biosynthesis; D006801:Humans; D013058:Mass Spectrometry; D012709:Serum Albumin / Q000737:chemistry* / Q000276:immunology* / Q000502:physiology; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D017127:Glycation End Products, Advanced; D012709:Serum Albumin