Abstract
Manual and automated solvent-free mini-ball mill (MBM) matrix-assisted laser desorption/ionization (MALDI) analysis of mixtures of β-amyloid peptides (1–11), (33–42), (1–42) and non-β-amyloid component of Alzheimer’s disease peptide yielded interpretable spectra for all of the peptides present regardless of their relative amounts in the samples. This was not the case for solvent-based MALDI analysis using traditional acidic aqueous/organic solvent conditions, which resulted in severe over-representation of hydrophilic peptide (1–11) and provided no spectra for insoluble amphiphilic peptide (1–42) even when present at 50% relative molar amount. Less accurate representation of components in mixtures by the traditional method appears to be a combination of poor dissolution of peptides in the solvent and preferential ionization of more hydrophilic peptides in the mixture. Consequently, only MBM provided a complete tryptic map of β-amyloid (1–42) compared to 67% coverage by traditional MALDI. Acetonitrile (0.1% TFA) led to improved coverage only at a 50% molar ratio of peptide (1–42), but also to a side product of (1–42), Met oxidation (amino acid 35), a phenomenon not observed in MBM MALDI analysis. Traditional MALDI analysis resulted in over-representation of hydrophilic soluble β-amyloid (1–11) in defined mixtures and autoproteolytic peptides of trypsin. In contrast, over-representation and under-representation were less pronounced in solvent-free MALDI in all of the investigated cases. Analysis of defined peptide and tryptic peptide mixtures showed that MBM MALDI yielded greater qualitative reliability, which also improved quantitative response relative to the solvent-based approach.
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Przybilla, L. M.; Brand, J. D.; Yoshimura, K.; Räder, H. J.; Müllen, K. Characterization of Large Synthetic Polycyclic Aromatic Hydrocarbons by MALDI- and LD-TOF Mass Spectrometry. Anal. Chem. 2000, 72, 4591–4597.
Trimpin, S.; Grimsdale, A. C.; Räder, H. J.; Müllen, K. Characterization of an Insoluble Poly(9,9-Diphenyl-2,7-Fluorene) by Solvent-Free Sample Preparation for MALDI-TOF Mass Spectrometry. Anal. Chem. 2002, 74, 3777–3782.
Trimpin, S.; Keune, S.; Räder, H. J.; Müllen, K. Solvent-free MALDI-MS: Developmental Improvements in the Reliability and the Potential of MALDI Analysis of Synthetic Polymers and Giant Organic Molecules. J. Am. Mass Spectrom. 2006, 17, 661–671.
Trimpin, S.; Deinzer, M. L. Solvent-free MALDI-MS for the Analysis of Biological Samples Via a Mini-Ball Mill Approach. J. Am. Mass Spectrom. 2005, 16, 542–547.
Turner, P. R.; O’Connor, K.; Tate, W. P.; Abraham, W. C. Roles of Amyloid Precursor Protein and Its Fragments in Regulating Neural Activity, Plasticity, and Memory. Prog. Neurobiol. 2003, 70, 1–32.
Soderberg, L.; Dahlqvist, C.; Kakuyama, H.; Thyberg, J.; Ito, A.; Winblad, B.; Naslund, J.; Tjernberg, L. O. Collagenous Alzheimer Amyloid Plaque Component Assembles Amyloid Fibrils Into Protease Resistant Aggregates. FEBS J 2005, 272(9), 2231–2236.
Tsubuki, S.; Takaki, Y.; Saido, T. C. Dutch, Flemish, Italian, and Arctic Mutations of APP and Resistance of Abeta to Physiologically Relevant Proteolytic Degradation. Lancet 2003, 361(9373), 1957–1958.
Perkins, D. N.; Pappin, D. J.; Creasy, D. M.; Cottrell, J. S. Probability-Based Protein Identification by Searching Sequence Databases Using Mass Spectrometry Data. Electrophoresis 1999, 18, 3551–3567.
Redeby, T.; Emmer, A. Membrane Protein and Peptide Sample Handling for MS Analysis Using a Structured MALDI Target. Anal. Bioanal. Chem. 2005, 381(1), 225–232.
Tummala, R.; Green-Church, K. B.; Limbach, P. A. Interactions Between Sodium Dodecyl Sulfate Micelles and Peptides During Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) of Proteolytic Digests. J. Am. Soc. Mass Spectrom. 2005, 16, 1438–1446.
Zischka, H.; Gloeckner, C. J.; Klein, C.; Willmann, S.; Swiatek-de Lange, M.; Ueffing, M. Improved Mass Spectrometric Identification of Gel-Separated Hydrophobic Membrane Proteins After Sodium Dodecyl Sulfate Removal by Ion-Pair Extraction. Proteomics 2004, 4(12), 3776–3782.
Zhang, N.; Li, L. Liquid Chromatography MALDI MS/MS for Membrane Proteome Analysis. Rapid Commun. Mass Spectrom. 2004, 18(8), 889–896.
Kheterpal, I.; Williams, A.; Murphy, C.; Bledsoe, B.; Wetzel, R. Structural Features of the Abeta Amyloid Fibril Elucidated by Limited Proteolysis. Biochemistry 2001, 40(39), 11757–11767.
Wang, M. Z.; Fitzgerald, M. C. A Solid Sample Preparation Method That Reduces Signal Suppression Effects in the MALDI Analysis of Peptides. Anal. Chem. 2001, 73, 625–631.
Liu, R.; Barkhordarian, H.; Emadi, S.; Park, C. B.; Sierks, M. R. Trehalose Differentially Inhibits Aggregation and Neurotoxicity of β-Amyloid 40 and 42. Neurobiol. Dis. 2005, 20(1), 74–81.
Malavolta, L.; Pinto, M. R.; Cuvero, J. H.; Nakaie, C. R. Interpretation of the Dissolution of Insoluble Peptide Sequences Based on the Acid-Base Properties of the Solvent. Protein Sci. 2006, 15(6), 1476–1488.
Speicher, K. D.; Kolbas, O.; Harper, S.; Speicher, D. W. Systematic Analysis of Peptide Recoveries from In-Gel Digestions for Protein Identifications in Proteome Studies. J. Biomol. Technol. 2000, 11, 74–86.
Stewart, I. I.; Thomson, T.; Figeys, D. 18O Labeling: A Tool for Proteomics. Rapid Commun. Mass Spectrom. 2001, 15, 2456–2465.
Trimpin, S.; Deinzer, M. L.. Anal. Chem. 2007, 1, 79(1), 71–78.
Wallace, W. E. Recent Advances in Quantitative Synthetic-Polymer Mass Spectrometry at NIST. Proceedings of the 52nd ASMS Conference on Mass Spectrometry and Allied Topics; Nashville, TN, May 2004.
Lalowski, M.; Golabek, A.; Lemere, C. A.; Selkoe, D. J.; Wisniewski, H. M.; Beavis, R. C.; Frangione, B.; Wisniewski, T. The “Nonamyloidogenic” p3 Fragment (Amyloid β 17–42) is a major constituent of Down’s Syndrome Cerebellar Pre-Amyloid. J. Biol. Chem. 1996, 271, 33623–33631.
Stapels, M. D.; Barofsky, D. F. Complementary Use of MALDI and ESI for the HPLC-MS/MS Analysis of DNA-Binding Proteins. Anal. Chem. 2004, 76, 5423–5430.
Bigwarfe, P. M. Jr.; Wood, T. D. Effect of Ionization Mode in the Analysis of Proteolytic Protein Digests. Int. J. Mass Spectrom. 2004, 234, 185–202.
Tummala, R.; Limbach, P. A. Serum Protein Profiling Using Surfactant-Aided Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. Anal. Chim. Acta 2005, 551, 137–141.
Butterfield, D. A.; Boyd-Kimball, D. The Critical Role of Methionine 35 in Alzheimer Amyloid β-Peptide (1–42)-Induced Oxidative Stress and Neurotoxicity. Biochim. Biophys. Acta 2005, 1703, 149–156.
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Published online April 29, 2007
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Trimpin, S., Deinzer, M.L. Solvent-free MALDI-MS for the analysis of β-amyloid peptides via the mini-ball mill approach: Qualitative and quantitative advances. J Am Soc Mass Spectrom 18, 1533–1543 (2007). https://doi.org/10.1016/j.jasms.2007.04.017
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DOI: https://doi.org/10.1016/j.jasms.2007.04.017