Analytical and Bioanalytical Chemistry

, Volume 389, Issue 4, pp 1075–1085

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

  • Yu Bai
  • Dmitry Galetskiy
  • Eugen Damoc
  • Jan Ripper
  • Markus Woischnik
  • Matthias Griese
  • Zhiqiang Liu
  • Shuying Liu
  • Michael Przybylski
Original Paper

DOI: 10.1007/s00216-007-1403-z

Cite this article as:
Bai, Y., Galetskiy, D., Damoc, E. et al. Anal Bioanal Chem (2007) 389: 1075. doi:10.1007/s00216-007-1403-z

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.

Keywords

Pulmonary alveolar proteinosis Lung surfactant proteins Bronchoalveolar lavage fluid Fourier transform ion cyclotron resonance mass spectrometry Proteome analysis 

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

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Yu Bai
    • 1
    • 2
  • Dmitry Galetskiy
    • 1
  • Eugen Damoc
    • 1
  • Jan Ripper
    • 3
  • Markus Woischnik
    • 3
  • Matthias Griese
    • 3
  • Zhiqiang Liu
    • 2
  • Shuying Liu
    • 2
  • Michael Przybylski
    • 1
  1. 1.Laboratory of Analytical Chemistry and Biopolymer Structure Analysis, Department of ChemistryUniversity of ConstanceConstanceGermany
  2. 2.Laboratory of New Drugs, Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchunChina
  3. 3.Department of Pediatric Pneumology, Dr. von Haunersches KinderspitalUniversity of MunichMunichGermany