Carbonylation of mitochondrial aconitase with 4-hydroxy-2-(E)-nonenal: Localization and relative reactivity of addition sites

4-Hydroxy-2-(E)-nonenal addition to porcine mitochondrial aconitase was investigated using mass spectrometry. Identified Michael addition sites are in red; underscores indicate sites detected with both ion trap and Orbitrap instruments (highest confidence). 94.2% sequence coverage was achieved using trypsin and chymotrypsin (trypsin only highlighted blue, chymotrypsin only highlighted yellow, and both enzymes highlighted green).469