Journal article
Ion Mobility-Mass Correlation Trend Line Separation of Glycoprotein Digests without Deglycosylation
International journal for ion mobility spectrometry, Vol.16(2), pp.105-115
06/01/2013
Handle:
https://hdl.handle.net/2376/105414
PMCID: PMC3728182
PMID: 23914139
Abstract
A high-throughput ion mobility mass spectrometer (IMMS) was used to rapidly separate and analyze peptides and glycopeptides derived from glycoproteins. Two glycoproteins, human α-1-acid glycoprotein and antithrombin III were digested with trypsin and subjected to electrospray traveling wave IMMS analysis. No deglycosylation steps were performed; samples were complex mixtures of peptides and glycopeptides. Peptides and glycosylated peptides with different charge states (up to 4 charges) were observed and fell on distinguishable trend lines in 2-D IMMS spectra in both positive and negative modes. The trend line separation patterns matched between both modes. Peptide sequence was identified based on the corresponding extracted mass spectra and collision induced dissociated (CID) experiments were performed for selected compounds to prove class identification. The signal-to-noise ratio of the glycopeptides was increased dramatically with ion mobility trend line separation compared to non-trend line separation, primarily due to selection of precursor ion subsets within specific mobility windows. In addition, isomeric mobility peaks were detected for specific glycopeptides. IMMS demonstrated unique capabilities and advantages for investigating and separating glycoprotein digests in this study and suggests a novel strategy for rapid glycoproteomics studies in the future.
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Details
- Title
- Ion Mobility-Mass Correlation Trend Line Separation of Glycoprotein Digests without Deglycosylation
- Creators
- Hongli Li - Department of Chemistry, Washington State University, Pullman, Washington, USBrad Bendiak - Department of Cell and Developmental Biology, Program in Structural Biology and Biophysics, University of Colorado, Health Sciences Center, Anschutz Medical Campus, Aurora, Colorado, USWilliam F Siems - Department of Chemistry, Washington State University, Pullman, Washington, USDavid R Gang - Institute of Biological Chemistry, Washington State University, Pullman, Washington, USHerbert H Hill - Department of Chemistry, Washington State University, Pullman, Washington, US
- Publication Details
- International journal for ion mobility spectrometry, Vol.16(2), pp.105-115
- Academic Unit
- Chemistry, Department of; Biological Chemistry, Institute of
- Grant note
- R33 RR020046 || RR / National Center for Research Resources : NCRR
- Identifiers
- 99900546828201842
- Language
- English
- Resource Type
- Journal article