期刊
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
卷 25, 期 12, 页码 2009-2019出版社
SPRINGER
DOI: 10.1007/s13361-014-0920-1
关键词
Ion mobility MS; Collision cross section; Traveling wave; CCS calibration; Gas-phase; Conformation; Proteomics; LysN
资金
- National Science Foundation [CHE-0957784]
- National Institutes of Health [1R01DK071801, 1R56DK071801]
- NIH [T32-GM008505]
- NSF Graduate Research Fellowship [DGE-1256259]
- H. I. Romnes Faculty Research Fellowship
Ion mobility (IM) is a gas-phase electrophoretic method that separates ions according to charge and ion-neutral collision cross-section (CCS). Herein, we attempt to apply a traveling wave (TW) IM polyalanine calibration method to shotgun proteomics and create a large peptide CCS database. Mass spectrometry methods that utilize IM, such as HDMSE, often use high transmission voltages for sensitive analysis. However, polyalanine calibration has only been demonstrated with low voltage transmission used to prevent gas-phase activation. If polyalanine ions change conformation under higher transmission voltages used for HDMSE, the calibration may no longer be valid. Thus, we aimed to characterize the accuracy of calibration and CCS measurement under high transmission voltages on a TW IM instrument using the polyalanine calibration method and found that the additional error was not significant. We also evaluated the potential error introduced by liquid chromatography (LC)-HDMSE analysis, and found it to be insignificant as well, validating the calibration method. Finally, we demonstrated the utility of building a large-population peptide CCS database by investigating the effects of terminal lysine position, via LysC or LysN digestion, on the formation of two structural sub-families formed by triply charged ions.
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