From Spectroglyph, LLC
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Nov 15, 2013: Editor's Highlights - Analytical Chemistry
- From Protein Complexes to Subunit Backbone Fragments: A Multistage Approach to Native Mass Spectrometry by Mikhail E. Belov,†,§ Eugen Damoc,† Eduard Denisov,† Philip D. Compton,‡ Stevan Horning,†Alexander A. Makarov,*,† and Neil L. Kelleher*,‡
Native mass spectrometry (MS) is becoming an important integral part of structural proteomics and system biology research. The approach holds great promise for elucidating higher levels of protein structure: from primary to quaternary. This requires the most efficient use of tandem MS, which is the cornerstone of MS-based approaches. In this work, we advance a two-step fragmentation approach, or (pseudo)-MS3, from native protein complexes to a set of constituent fragment ions. Using an efficient desolvation approach and quadrupole selection in the extended mass-to-charge (m/z) range, we have accomplished sequential dissociation of large protein complexes, such as phosporylase B (194kDa), pyruvate kinase (232 kDa), and GroEL (801 kDa), to highlycharged monomers which were then dissociated to a set of multiply charged fragmentation products. Fragment ion signals were acquired with a high resolution, high mass accuracy Orbitrap instrument that enabled highly confident identifications of the precursor monomer subunits. The developed approach is expected to enable characterization of stoichiometry and composition of endogenous native protein complexes at an unprecedented level of detail.
May 1, 2013: Spectroglyph has been awarded NIH grant
Spectroglyph, LLC received a grant, ref 1R43GM105152-01A1, from the National Institutes of Health to develop a Novel Hybrid Bioanalytical System.
Application Notes:
- MALDI ESI Injector
- Bridging the Gap between Ion Mobility Spectrometry and an Orbitrap
- Extending the upper mass limit of Orbitrap Mass Spectrometry
- Increasing Efficiency of Surface Acoustic Wave Nebulization (SAWN) with an Ion Funnel
- IFT Injector Interface
- Bottom-up proteomics with HeLa cell digests
- From Protein Complexes to Subunit Backbone Fragments: A Multistage Approach to Native Mass Spectrometry
- Multi-step Sequencing and Confident Identification of Native Protein Complexes on an Orbitrap Mass Spectrometer
- Triple-Stage Mass Spectrometry Unravels the Heterogeneity of an Endogenous Protein Complex
- Design and Performance of a Novel Interface for Combined MALDI at Elevated Pressure and ESI with Orbitrap Mass Spectrometry
- Laser post-ionization combined with a high resolving power Orbitrap for enhanced MALDI-MS imaging of lipids
- MALDI/ESI Injector Data
- MALDI/ESI Injector User Manual