title: Electrocatalytic assay for monitoring methylglyoxal-mediated protein glycation.
abstract: Protein glycation is a complex process that plays an important role in diabetes mellitus, aging, and the regulation of protein function in general. As a result, current methodological research on proteins is focused on the development of novel approaches for investigating glycation and the possibility of monitoring its modulation and selective inhibition. In this paper, a first sensing strategy for protein glycation is proposed, based on protein electroactivity measurement. Concretely, the label-free method proposed is based on the application of a constant-current chronopotentiometric stripping (CPS) analysis at Hg-containing electrodes. The glycation process was monitored as the decrease in the electrocatalytic protein signal, peak H, observed at highly negative potentials at around -1.8 V (vs Ag/AgCl3 M KCl), which was previously ascribed to a catalytic hydrogen evolution reaction (CHER). Using this method, a model protein bovine serum albumin was investigated over 3 days of incubation with the glycation agent methylglyoxal in the absence or presence of the glycation inhibitor aminoguanidine (pimagedine). The electrochemical methodology presented here could open up new possibilities in research on protein glycation and oxidative modification. The methodology developed also provides a new option for the analysis of protein intermolecular interactions using electrochemical sensors, which was demonstrated by the application of a silver solid amalgam electrode (AgSAE) for monitoring the glycation process in samples of bovine serum albumin, human serum albumin, and lysozyme.
authors.affiliation: Department of Medical Chemistry and Biochemistry, Faculty of Medicine and Dentistry, Palacky University , Hnevotinska 3, 775 15 Olomouc, Czech Republic.
authors.forename: Marika
authors.initials: M
authors.lastname: Havlikova
authors.forename: Martina
authors.initials: M
authors.lastname: Zatloukalova
authors.forename: Jitka
authors.initials: J
authors.lastname: Ulrichova
authors.forename: Petr
authors.initials: P
authors.lastname: Dobes
authors.forename: Jan
authors.initials: J
authors.lastname: Vacek
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D002384:Catalysis; D002417:Cattle; D004563:Electrochemistry; D004566:Electrodes; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017127:Glycation End Products, Advanced; D006031:Glycosylation / Q000187:drug effects*; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009113:Muramidase / Q000032:analysis* / Q000737:chemistry; D011487:Protein Conformation; D011765:Pyruvaldehyde / Q000494:pharmacology*; D012709:Serum Albumin / Q000032:analysis* / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000737:chemistry; D000093423:Glycated Serum Albumin
chemical_list: D004791:Enzyme Inhibitors; D017127:Glycation End Products, Advanced; D006146:Guanidines; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D011765:Pyruvaldehyde; D009113:Muramidase; C004479:pimagedine; D000093423:Glycated Serum Albumin