ArticleJournal of the American Society for Mass Spectrometry2010
Resolving the composition of protein complexes using a MALDI LTQ Orbitrap.
Article in Journal of the American Society for Mass Spectrometry, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
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- Astrochemical Pathways to Complex Organic and Prebiotic Molecules: Experimental Perspectives for In Situ Solid-State Studies.Life (Basel, Switzerland) · 2021Review
- Interactome and Proteome Dynamics Uncover Immune Modulatory Associations of the Pathogen Sensing Factor cGAS.Cell systems · 2018Article
- The functional interactome of PYHIN immune regulators reveals IFIX is a sensor of viral DNA.Molecular systems biology · 2015Article
- A proteomic perspective of Sirtuin 6 (SIRT6) phosphorylation and interactions and their dependence on its catalytic activity.Molecular & cellular proteomics : MCP · 2014Article
- Proteomics-based methods for discovery, quantification, and validation of protein-protein interactions.Analytical chemistry · 2013Review
- A proteomic approach provides new insights into the control of soil-borne plant pathogens by Bacillus species.PloS one · 2013Article
- Evidence for a genetic and physical interaction between nonstructural proteins NS1 and NS4B that modulates replication of West Nile virus.Journal of virology · 2012Article
- Acetylation modulates cellular distribution and DNA sensing ability of interferon-inducible protein IFI16.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Proteomic characterization of pseudorabies virus extracellular virions.Journal of virology · 2011Article
- Identification of staphylococcal nuclease domain-containing 1 (SND1) as a Metadherin-interacting protein with metastasis-promoting functions.The Journal of biological chemistry · 2011Article
- A conserved coatomer-related complex containing Sec13 and Seh1 dynamically associates with the vacuole in Saccharomyces cerevisiae.Molecular & cellular proteomics : MCP · 2011Article
- The free and bound forms of Lpp occupy distinct subcellular locations in Escherichia coli.Molecular microbiology · 2011Article
- Nuclear import of histone deacetylase 5 by requisite nuclear localization signal phosphorylation.Molecular & cellular proteomics : MCP · 2011Article
- Human cytomegalovirus pUL83 stimulates activity of the viral immediate-early promoter through its interaction with the cellular IFI16 protein.Journal of virology · 2010Article
- The response regulator SprE (RssB) is required for maintaining poly(A) polymerase I-degradosome association during stationary phase.Journal of bacteriology · 2010Article
- Host factors associated with the Sindbis virus RNA-dependent RNA polymerase: role for G3BP1 and G3BP2 in virus replication.Journal of virology · 2010Article
- Human cytomegalovirus UL29/28 protein interacts with components of the NuRD complex which promote accumulation of immediate-early RNA.PLoS pathogens · 2010Article
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Authors and funding
5 authors.
Funding
Abstract
Current biological studies have been advanced by the continuous development of robust, accurate, and sensitive mass spectrometric technologies. The MALDI LTQ Orbitrap is a new addition to the Orbitrap configurations, known for their high resolving power and accuracy. This configuration provides features inherent to the MALDI source, such as reduced spectra complexity, forgiveness to contaminants, and sample retention for follow-up analyses with targeted or hypothesis-driven questions. Here we investigate its performance for characterizing the composition of isolated protein complexes. To facilitate the assessment, we selected two well characterized complexes from Saccharomyces cerevisiae, Apl1 and Nup84. Manual and automatic MS and MS/MS analyses readily resolved their compositions, with increased confidence of protein identification compared with our previous reports using MALDI QqTOF and MALDI IT. CID fragmentation of singly-charged peptides provided sufficient information for conclusive identification of the isolated proteins. We then assessed the resolution, accuracy, and sensitivity provided by this instrument in the context of analyzing the isolated protein assemblies. Our analysis of complex mixtures of singly-charged ions up to m/z 4000 showed that (1) the resolving power, inversely proportional to the square root of m/z, had over four orders of magnitude dynamic range; (2) internal calibration led to improved accuracy, with an average absolute mass error of 0.5 ppm and a distribution centered at 0 ppm; and (3) subfemtomole sensitivity was achieved using both CHCA and DHB matrices. Additionally, our analyses of a synthetic phosphorylated peptide in mixtures showed subfemtomole level of detection using neutral loss scanning.
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