Analytical and bioanalytical chemistry, 2020-07, Vol.412 (18), p.4245-4259
2020
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- Autor(en) / Beteiligte
- Titel
- Unveiling the molecular mechanisms underpinning biorecognition of early-glycated human serum albumin and receptor for advanced glycation end products
- Ist Teil von
- Analytical and bioanalytical chemistry, 2020-07, Vol.412 (18), p.4245-4259
- Ort / Verlag
- Berlin/Heidelberg: Springer Berlin Heidelberg
- Erscheinungsjahr
- 2020
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Serum levels of early-glycated albumin are significantly increased in patients with diabetes mellitus and may play a role in worsening inflammatory status and sustaining diabetes-related complications. To investigate possible pathological recognition involving early-glycated albumin and the receptor for advanced glycation end products (RAGE), an early-glycated human serum albumin (HSAgly), with a glycation pattern representative of the glycated HSA form abundant in diabetic patients, and the recombinant human RAGE ectodomain (VC1) were used. Biorecognition between the two interactants was investigated by combining surface plasmon resonance (SPR) analysis and affinity chromatography coupled with mass spectrometry (affinity-MS) for peptide extraction and identification. SPR analysis proved early-glycated albumin could interact with the RAGE ectodomain with a steady-state affinity constant of 6.05 ± 0.96 × 10 −7 M. Such interaction was shown to be specific, as confirmed by a displacement assay with chondroitin sulfate, a known RAGE binder. Affinity-MS studies were performed to map the surface area involved in the recognition. These studies highlighted that a region surrounding Lys525 and part of subdomain IA were involved in VC1 recognition. Finally, an in silico analysis highlighted (i) a key role for glycation at Lys525 (the most commonly glycated residue in HSA in diabetic patients) through a triggering mechanism similar to that previously observed for AGEs or advanced lipoxidation end products and (ii) a stabilizing role for subdomain IA. Albeit a moderate affinity for complex formation, the high plasma levels of early-glycated albumin and high percentage of glycation at Lys525 in diabetic patients make this interaction of possible pathological relevance. Graphical abstract
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1618-2642eISSN: 1618-2650DOI: 10.1007/s00216-020-02674-wTitel-ID: cdi_proquest_miscellaneous_2398625845
- Format
- –
- Schlagworte
- Advanced glycosylation end products, Affinity, Affinity chromatography, Albumin, Analytical Chemistry, Binding Sites, Biochemistry, Characterization and Evaluation of Materials, Chemistry, Chemistry and Materials Science, Chondroitin sulfate, Chromatography, Affinity, Complex formation, Diabetes, Diabetes mellitus, Diabetes Mellitus - metabolism, Diabetics, Ethylenediaminetetraacetic acid, Food Science, Glycosylation, Human serum albumin, Humans, Inflammation, Laboratory Medicine, Lipid peroxidation, Mass spectrometry, Mass spectroscopy, Models, Molecular, Molecular modelling, Monitoring/Environmental Analysis, Paper in Forefront, Plasma levels, Protein Binding, Receptor for Advanced Glycation End Products - chemistry, Receptor for Advanced Glycation End Products - metabolism, Receptors, Recognition, Recombinant Proteins - chemistry, Recombinant Proteins - metabolism, Serum albumin, Serum Albumin - chemistry, Serum Albumin - metabolism, Serum Albumin, Human - chemistry, Serum Albumin, Human - metabolism, Serum levels, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Surface Plasmon Resonance