Stability study of Rhodobacter capsulatus ferrocytochrome c 2 wild‐type and site‐directed mutants using hydrogen/deuterium exchange monitored by electrospray ionization mass spectrometry

摘要

>To estimate the stability of Rhodobacter capsulatus ferrocytochrome c ₂ wild-type and site-directed mutants, charge state distributions and hydrogen/deuterium exchange rates were monitored by electrospray ionization mass spectrometry. The relative stability of the mutants was observed with the order: V11 insert Y75F wild-type = K32E K12D = K14E ≥ K52E K14E/K32E W67Y P35A 157N G34S. (A preliminary account has been presented for mutants G34S and P35A [Jaquinod et al. (1995) Rapid Commun. Mass Spectrom. 9, 1135–1140].) This approach is shown to be a useful tool for rapid characterization of mutational effects on protein conformation.