muramidase and glycolaldehyde

Glycation occurs in vivo as a result of the nonenzymatic reaction of carbohydrates (and/or their autoxidation products) with proteins, DNA, or lipids. Protein glycation causes loss-of-function and, consequently, the development of diabetic-related diseases. Glycation also boosts protein aggregation, which can be directly related with the higher prevalence of aggregating diseases in diabetic people. However, the molecular mechanism connecting glycation with aggregation still remains unclear. Previously we described mechanistically how glycation of hen egg-white lysozyme (HEWL) with ribose induced its aggregation. Here we address the question of whether the ribose-induced aggregation is a general process or it depends on the chemical nature of the glycating agent. Glycation of HEWL with glycolaldehyde occurs through two different scenarios depending on the HEWL concentration regime (both within the micromolar range). At low HEWL concentration, non-cross-linking fluorescent advanced glycation end-products (AGEs) are formed on Lys side chains, which do not change the protein structure but inhibit its enzymatic activity. These AGEs have little impact on HEWL surface hydrophobicity and, therefore, a negligible effect on its aggregation propensity. Upon increasing HEWL concentration, the glycation mechanism shifts toward the formation of intermolecular cross-links, which triggers a polymerization cascade involving the formation of insoluble spherical-like aggregates. These results notably differ with the aggregation-modulation mechanism of ribosylated HEWL directed by hydrophobic interactions. Additionally, their comparison constitutes the first experimental evidence showing that the mechanism underlying the aggregation of a glycated protein depends on the chemical nature of the glycating agent.

Topics: Acetaldehyde; Animals; Chickens; Diabetes Mellitus; Glycation End Products, Advanced; Glycosylation; Hydrophobic and Hydrophilic Interactions; Muramidase; Protein Conformation; Proteins; Spectrometry, Fluorescence; Surface Properties

Innate immunity directs the adaptive immune response by identifying antigens that are associated with infectious agents. Although some microbial antigens can be recognized by innate immune receptors, most cannot, and these require identification by some other means. The introduction of aldehydes into antigens by glycolaldehyde, which can be produced by activated neutrophils reacting with serine, or by the oxidation of an N-linked oligosaccharide with NaIO4, enhances by several orders of magnitude their immunogenicity in mice. The augmented immunogenicity requires the presence of an aldehyde on the antigen, and is not dependent on protein aggregation. An in vitro correlate of augmented immunogenicity is the enhanced presentation of glycolaldehyde-modified antigen to T cells by macrophages and bone marrow-derived dendritic cells. The potential clinical importance of this form of antigen modification is twofold: glycolaldehyde renders a model self antigen immunogenic, and it converts a relatively non-immunogenic malaria antigen, merozoite surface protein-1, into an effective immunogen. Thus, the tagging of antigens by the addition of aldehydes, which may be an innate immune mechanism to facilitate their recognition by the adaptive immune system, may have a role in the genesis of autoimmunity and the development of vaccines.

Topics: Acetaldehyde; Aldehydes; Animals; Antibody Formation; Antigens; Antigens, Protozoan; Autoantigens; Autoimmunity; Chickens; Columbidae; Cytochrome c Group; Immunity, Innate; Merozoite Surface Protein 1; Mice; Muramidase; Neutrophils; Ovalbumin; Oxidation-Reduction; Periodic Acid; Serine; Structure-Activity Relationship; Vaccines

Horseradish peroxidase (EC 1.11.1.7) is shown to catalyze the aerobic oxidation of lysozyme, bovine serum albumin, and protamine adducts with glycolaldehyde at physiological pH. This reaction is accompanied by light emission, which is attributed to the generation of triplet species. The intensity of chemiluminescence is enhanced by addition of chlorophyll alpha solubilized in Brij 35. A role of electronically excited species in deleterious and pathological processes associated with formation of Schiff-type adducts is suggested, with emphasis on the case of alcohol-induced liver injury.

Topics: Acetaldehyde; Chlorophyll; Chlorophyll A; Horseradish Peroxidase; Kinetics; Luminescent Measurements; Muramidase; Oxidation-Reduction; Oxygen; Peroxidases; Polidocanol; Polyethylene Glycols; Protamines; Proteins; Serum Albumin, Bovine; Spectrophotometry