Effects of ion/ion proton transfer reactions on conformation of gas-phase cytochrome c ions.
Positive ions from cytochrome c are studied in a 3-D ion trap/ion mobility (IM)/quadrupole-time-of-flight (TOF) instrument with three independent ion sources. The IM separation allows measurement of the cross section of the ions. Ion/ion reactions in the 3-D ion trap that remove protons cause the cytochrome c ions to refold gently without other degradation of protein structure, i.e., fragmentation or loss of heme group or metal ion. The conformation(s) of the product ions generated by ion/ion reactions in a given charge state are similar regardless of whether the cytochrome c ions are originally in +8 or +9 charge states. In the lower charge states (+1 to +5) cytochrome c ions made by the ion/ion reaction yield a single IM peak with cross section of approximately 1110 to 1180 A(2), even if the original +8 ion started with multiple conformations. The conformation expands slightly when the charge state is reduced from +5 to +1. For product ions in the +6 to +8 charge states, ions created from higher charge states (+9 to +16) by ion/ion reaction produce more compact conformation(s) in somewhat higher abundances compared with those produced directly by the electrospray ionization (ESI) source. For ions in intermediate charge states that have a variety of resolvable conformers, the voltage used to inject the ions into the drift tube, and the voltage and duration of the pulse that extracts ions from the ion trap, can affect the observed abundances of various conformers.
Zhao
Qin
Q
Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
Schieffer
Gregg M
GM
Soyk
Matthew W
MW
Anderson
Timothy J
TJ
Houk
R S
RS
Badman
Ethan R
ER
D000818:Animals; D002417:Cattle; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism; D005740:Gases / Q000737:chemistry; D007477:Ions / Q000737:chemistry* / Q000378:metabolism; D000432:Methanol / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D011522:Protons; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D005740:Gases; D007477:Ions; D011522:Protons; D045304:Cytochromes c; D000432:Methanol