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High resolution mass spectrometric alveolar proteomics: Identification of surfactant protein SP‐A and SP‐D modifications in proteinosis and cystic fibrosis patients
Author(s) -
Bai Yu,
Galetskiy Dmitry,
Damoc Eugen,
Paschen Christian,
Liu Zhiqiang,
Griese Mathias,
Liu Shuying,
Przybylski Michael
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200400855
Subject(s) - chemistry , mass spectrometry , proteome , proteomics , chromatography , fourier transform ion cyclotron resonance , pulmonary alveolar proteinosis , pulmonary surfactant , gel electrophoresis , biochemistry , lung , medicine , gene
In the present study, one‐ and two‐dimensional gel electrophoresis combined with high resolution Fourier transform‐ion cyclotron resonance mass spectrometry (FT‐ICR MS) have been applied as powerful approaches for the proteome analysis of surfactant proteins SP‐A and SP‐D, including identification of structurally modified and truncation forms, in bronchoalveolar lavage fluid from patients with cystic fibrosis, chronic bronchitis and pulmonary alveolar proteinosis. Highly sensitive micropreparation techniques were developed for matrix‐assisted laser desorption/ionization (MALDI) FT‐ICR MS analysis which provided the identification of surfactant proteins at very low levels. Owing to the high resolution, FT‐ICR MS was found to provide substantial advantages for the structural identification of surfactant proteins from complex biological matrices with high mass determination accuracy. Several protein bands corresponding to SP‐A and SP‐D were identified by MALDI‐FT‐ICR MS after electrophoretic separation by one‐ and two‐dimensional gel electrophoresis, and provided the identification of structural modifications (hydroxy‐proline) and degradation products. The high resolution mass spectrometric proteome analysis should facilitate the unequivocal identification of subunits, aggregations, modifications and degradation products of surfactant proteins and hence contribute to the understanding of the mechanistic basis of lung disease pathogenesis.