This isolation strategy, coupled
with LC–MS–MS, allowed the identification of 417 discernible proteins
with a statistically significant increase in adduction with increasing HNE
exposure [44]. Western blot analysis of a subset of the streptavidin captured
proteins showed adduction at HNE concentrations as low as 1 μM.
Thus, the sensitivity of the biotin hydrazide–streptavidin capture and release
approach allows a global analysis of electrophile-adducted proteins
at physiologically relevant concentrations of HNE. Furthermore, protein
interaction network analysis of the proteomics data showed enrichment
in several important subsystems following HNE treatment, including oxidative
stress response, proteasomal degradation, protein folding, and ribonucleoproteins,
among others [44]. This study highlights the relevance
of capture and release approaches involving biotin and streptavidinwith
respect to gaining a global perspective of cellular systems impacted by
lipid electrophile adduction of proteins.