Abstract
Correct charge state assignment is crucial to assigning an accurate mass to supramolecular complexes analyzed by electrospray mass spectrometry. Conventional charge state assignment techniques fall short of reliably and unambiguously predicting the correct charge state for many supramolecular complexes. We provide an explanation of the shortcomings of the conventional techniques and have developed a robust charge state assignment method that is applicable to all spectra.
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Ganem, B.; Li, Y. T.; Henion, J. D. Observation of Noncovalent Enzyme Substrate and Enzyme Product Complexes by Ion-Spray Mass-Spectrometry. J. Am. Chem. Soc. 1991, 11(20), 7818–7819.
Fitzgerald, M. C.; Chernushevich, I.; Standing, K. G.; Whitman, C. P.; Kent, S. B. H. Probing the Oligomeric Structure of an Enzyme by Electrospray Ionization Time-of-Flight Mass Spectrometry. Proc. Nat. Acad. Sci. U. S. A. 1996, 9(14), 6851–6856.
Loo, J. A. Studying Noncovalent Protein Complexes by Electrospray Ionization Mass Spectrometry. Mass Spectrom. Rev. 1997, 1(1), 1–23.
Rostom, A. A.; Fucini, P.; Benjamin, D. R.; Juenemann, R.; Nierhaus, K. H.; Hartl, F. U.; Dobson, C. M.; Robinson, C. V. Detection and Selective Dissociation of Intact Ribosomes in a Mass Spectrometer. Proc. Nat. Acad. Sci. U. S. A. 2000, 9(10), 5185–5190.
Tito, M. A.; Tars, K.; Valegard, K.; Hajdu, J.; Robinson, C. V. Electrospray Time-of-Flight Mass Spectrometry of the Intact MS2 Virus Capsid. J. Am. Chem. Soc. 2000, 12(14), 3550–3551.
Van Berkel, W. J. H.; Van Den Heuvel, R. H. H.; Versluis, C.; Heck, A. J. R. Detection of Intact Megadalton Protein Assemblies of Vanillyl-Alcohol Oxidase by Mass Spectrometry. Protein Sci. 2000, 9, 435–439.
Sanglier, S.; Leize, E.; Van Dorsselaer, A.; Zal, F. Comparative ESI-MS Study of Similar to 2. 2 MDa Native Hemocyanin from Deep-Sea and Shore Crabs: From Protein Oligomeric State to Biotope. J. Am. Soc. Mass Spectrom. 2003, 1(5), 419–429.
Green, B. N.; Bordoli, R. S.; Hanin, L. G.; Lallier, F. H.; Toulmond, A.; Vinogradov, S. N. Electrospray Ionization Mass Spectrometric Determination of the Molecular Mass of the Similar to 200-kDa Globin Dodecamer Subassemblies in Hexagonal Bilayer Hemoglobins. J. Biol. Chem. 1999, 27(40), 28206–28212.
Tang, X. J.; Brewer, C. F.; Saha, S.; Chernushevich, I.; Ens, W.; Standing, K. G. Investigation of Protein-Protein Noncovalent Interactions in Soybean Agglutinin by Electrospray-Ionization Time-of-Flight Mass-Spectrometry. Rapid Commun. Mass Spectrom. 1994, 8(9), 750–754.
Chernushevich, I. V.; Thomson, B. A. Collisional Cooling of Large Ions in Electrospray Mass Spectrometry. Anal. Chem. 2004, 7(6), 754–1760.
Smith, R. D.; Lightwahl, K. J.; Winger, B. E.; Loo, J. A. Preservation of Noncovalent Associations in Electrospray Ionization Mass-Spectrometry-Multiply Charged Polypeptide and Protein Dimers. Org. Mass Spectrom. 1992, 2, 811–821.
Siuzdak, G.; Bothner, B.; Yeager, M.; Brugidou, C.; Fauquet, C. M.; Hoey, K.; Chang, C. M. Mass Spectrometry and Viral Analysis. Chem. Biol. 1996, 3(1), 45–48.
Sobott, F.; Benesch, J. L. P.; Vierling, E.; Robinson, C. V. Subunit Exchange of Multimeric Protein Complexes—Real-Time Monitoring of Subunit Exchange Between Small Heat Shock Proteins by Using Electrospray Mass Spectrometry. J. Biol. Chem. 2002, 27(41), 38921–38929.
Benesch, J. L. P.; Sobott, F.; Robinson, C. V. Thermal Dissociation of Multimeric Protein Complexes by Using Nanoelectrospray Mass Spectrometry. Anal. Chem. 2003, 7(10), 2208–2214.
Ilag, L. L.; Videler, H.; McKay, A. R.; Sobott, F.; Fucini, P.; Nierhaus, K. H.; Robinson, C. V. Heptameric (L12)(6)/L10 Rather than Canonical Pentameric Complexes are Found by Tandem MS of Intact Ribosomes from Thermophilic Bacteria. Proc. Nat. Acad. Sci. U. S. A. 2005, 10(23), 8192–8197.
Taverner, T.; Hernandez, H.; Sharon, M.; Ruotolo, B. T.; Matak-Vinkovic, D.; Devos, D.; Russell, R. B.; Robinson, C. V. Subunit Architecture of Intact Protein Complexes from Mass Spectrometry and Homology Modeling. Acc. Chem. Res. 2008, 4(5), 617–627.
Sobott, F.; Robinson, C. V. Characterizing Electrosprayed Biomolecules Using Tandem-MS—the Noncovalent GroEL Chaperonin Assembly. Int. J. Mass Spectrom. 2004, 236(1/3), 25–32.
Mckay, A. R.; Ruotolo, B. T.; Ilag, L. L.; Robinson, C. V. Mass Measurements of Increased Accuracy Resolve Heterogeneous Populations of Intact Ribosomes. J. Am. Chem. Soc. 2006, 12(35), 11433–11442.
Allen, M.; Willits, D.; Mosolf, J.; Young, M.; Douglas, T. Protein Cage Constrained Synthesis of Ferrimagnetic Iron Oxide Nanoparticles. Adv. Mater. 2002, 1(21), 1562–1565.
Wiedenheft, B.; Mosolf, J.; Willits, D.; Yeager, M.; Dryden, K. A.; Young, M.; Douglas, T. An Archaeal Antioxidant: Characterization of a Dps-Like Protein from Sulfolobus solfataricus. Proc. Nat. Acad. Sci. U. S. A. 2005, 10(30), 10551–10556.
Fernandez De la; Mora, J. Electrospray Ionization of Large Multiply Charged Species Proceeds Via Dole’s Charged Residue Mechanism. Anal. Chim. Acta. 2000, 40(1), 93–104.
Heck, A. J. R.; van den Heuvel, R. H. H. Investigation of Intact Protein Complexes by Mass Spectrometry. Mass Spectrom. Rev. 2004, 2(5), 368–389.
Gauss, G. H.; Benas, P.; Wiedenheft, B.; Young, M.; Douglas, T.; Lawrence, C. M. Structure of the DPS-Like Protein from Sulfolobus solfataricus Reveals a Bacterioferritin-Like Dimetal Binding Site Within a DPS-Like Dodecameric Assembly. Biochemistry 2006, 45(36), 10815–10827.
van Breukelen, B. Resolving Stoichiometries and Oligomeric States of Glutamate Synthase Protein Complexes with Curve Fitting and Simulation of Electrospray Mass Spectra. Rapid Commun. Mass Spectrom. 2006, 2(16), 2490–2496.
Pinkse, M. W. H.; Maier, C. S.; Kim, J. I.; Oh, B. H.; Heck, A. J. R. Macromolecular Assembly of Helicobacter pylori Urease Investigated by Mass Spectrometry. J. Mass Spectrom. 2003, 3(3), 315–320.
Ilari, A.; Stefanini, S.; Chiancone, E.; Tsernoglou, D. The Dodecameric Ferritin from Listeria innocua Contains a Novel Intersubunit Iron-Binding Site. Nat. Struct. Biol. 2000, 7(1), 38–43.
Benesch, J. L. P.; Ruotolo, B. T.; Simmons, D. A.; Robinson, C. V. Protein Complexes in the Gas Phase: Technology for Structural Genomics and Proteomics. Chem. Rev. 2007, 10(8), 3544–3567.
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Published online November 17, 2008
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Liepold, L., Oltrogge, L.M., Suci, P.A. et al. Correct charge state assignment of native electrospray spectra of protein complexes. J Am Soc Mass Spectrom 20, 435–442 (2009). https://doi.org/10.1016/j.jasms.2008.10.024
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DOI: https://doi.org/10.1016/j.jasms.2008.10.024