Biophysical and mass spectrometry based characterization of methylglyoxal-modified myoglobin: Role of advanced glycation end products in inducing protein structural alterations

Methylglyoxal (MG) is a highly reactive alpha-dicarbonyl compound which reacts with proteins to form advanced glycation end products (AGEs). MG-induced AGE (MAGE) formation is particularly significant in diabetic condition. In the current study, we have undertaken a time-dependant characterization of MG-modified myoglobin following incubation of the heme protein with the alpha-dicarbonyl compound for different time periods. Interestingly, mass spectrometric studies indicated modifications at two specific lysine residues, Lys-87 and Lys-133. The AGE adducts identified at Lys-87 were carboxymethyllysine and carboxyethyllysine, while those detected at Lys133 included pyrraline-carboxymethyllysine and carboxyethyllysine, respectively. Far-UV CD studies revealed a decrease in the native alpha-helical content of the heme protein gradually with increasing time of MG incubation. In addition, MG modification was found to induce changes in tertiary structure as well as surface hydrophobicity of the heme protein. MG-derived AGE adducts thus appear to alter the structure of Mb considerably. Considering the increased level of MG in diabetic condition, the current study appears physiologically relevant in terms of understanding AGE-mediated protein modification and subsequent structural changes.

Automated summary

The study characterized MG-modified myoglobin in a time-dependent manner, with mass spectrometric studies indicating modifications at Lys-87 and Lys-133. Far-UV CD studies revealed alterations in protein structure and surface hydrophobicity as a result of MG modification. These findings suggest that MG-derived AGE adducts significantly impact the structure of Mb.