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Lung alveolar proteomics of bronchoalveolar lavage from a pulmonary alveolar proteinosis patient using high-resolution FTICR mass spectrometry

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Abstract

High-resolution Fourier transform ion cyclotron resonance (FTICR) mass spectrometry was developed and applied to the proteome analysis of bronchoalveolar lavage fluid (BALF) from a patient with pulmonary alveolar proteinosis. With use of 1-D and 2-D gel electrophoresis, surfactant protein A (SP-A) and other surfactant-related lung alveolar proteins were efficiently separated and identified by matrix-assisted laser desorption/ionization FTICR mass spectrometry . Low molecular mass BALF proteins were separated using a gradient 2-D gel. An efficient extraction/precipitation system was developed and used for the enrichment of surfactant proteins. The result of the BALF proteome analysis show the presence of several isoforms of SP-A, in which an N-non-glycosylierte form and several proline hydroxylations were identified. Furthermore, a number of protein spots were found to contain a mixture of proteins unresolved by 2-D gel electrophoresis, illustrating the feasibility of high-resolution mass spectrometry to provide identifications of proteins that remain unseparated in 2-D gels even upon extended pH gradients.

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Abbreviations

FTICR-MS:

Fourier transform ion cyclotron resonance mass spectrometry

BALF:

bronchoalveolar lavage fluid

PAP:

pulmonary alveolar proteinosis

GM-CSF:

granulocyte-macrophage colony stimulating factor

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Acknowledgements

Y.B. acknowledges a scholarship from the Max Planck Society. We gratefully acknowledge the expert assistance of Nikolay Youhnovski with FTICR-MS procedures. This work was supported by grants from the Deutsche Forschungsgemeinschaft (Alveolar Proteomics, to M.G. and M.P; and Biopolymer-MS, to M.P.) and the Fonds der Chemischen Industrie.

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Authors and Affiliations

  1. Laboratory of Analytical Chemistry and Biopolymer Structure Analysis, Department of Chemistry, University of Constance, Box M 731, 78457, Constance, Germany

    Yu Bai, Dmitry Galetskiy, Eugen Damoc & Michael Przybylski

  2. Laboratory of New Drugs, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 159 Remin Street, 130022, Changchun, China

    Yu Bai, Zhiqiang Liu & Shuying Liu

  3. Department of Pediatric Pneumology, Dr. von Haunersches Kinderspital, University of Munich, Lindwurmstr. 4, 80337, Munich, Germany

    Jan Ripper, Markus Woischnik & Matthias Griese

Authors
  1. Yu Bai
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  2. Dmitry Galetskiy
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  3. Eugen Damoc
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  4. Jan Ripper
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  5. Markus Woischnik
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  6. Matthias Griese
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  7. Zhiqiang Liu
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  8. Shuying Liu
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  9. Michael Przybylski
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Corresponding author

Correspondence to Michael Przybylski.

Additional information

Yu Bai and Dmitry Galetskiy both contributed equally to this work.

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Bai, Y., Galetskiy, D., Damoc, E. et al. Lung alveolar proteomics of bronchoalveolar lavage from a pulmonary alveolar proteinosis patient using high-resolution FTICR mass spectrometry. Anal Bioanal Chem 389, 1075–1085 (2007). https://doi.org/10.1007/s00216-007-1403-z

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  • DOI: https://doi.org/10.1007/s00216-007-1403-z

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