Stability study of Rhodobacter capsulatus ferrocytochrome c2 wild-type and site-directed mutants using hydrogen/deuterium exchange monitored by electrospray ionization mass spectrometry.
To estimate the stability of Rhodobacter capsulatus ferrocytochrome c2 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 > or = K52E > K14E/K32E > W67Y > P35A > I57N > 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.
Jaquinod
M
M
Institut de Biologie Structurale Jean-Pierre Ebel, CEA / CNRS, Grenoble, France.
Guy
P
P
Halgand
F
F
Caffrey
M
M
Fitch
J
J
Cusanovich
M
M
Forest
E
E
D003574:Cytochrome c Group / Q000737:chemistry* / Q000235:genetics; D003903:Deuterium / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011487:Protein Conformation; D016354:Rhodobacter capsulatus / Q000737:chemistry*
D003574:Cytochrome c Group; D006859:Hydrogen; D003903:Deuterium