Lung alveolar proteomics of bronchoalveolar lavage from a pulmonary alveolar proteinosis patient using high-resolution FTICR mass spectrometry.
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.
Bai
Yu
Y
Laboratory of Analytical Chemistry and Biopolymer Structure Analysis, Department of Chemistry, University of Constance, 78457, Constance, Germany.
Galetskiy
Dmitry
D
Damoc
Eugen
E
Ripper
Jan
J
Woischnik
Markus
M
Griese
Matthias
M
Liu
Zhiqiang
Z
Liu
Shuying
S
Przybylski
Michael
M
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D017357:Cyclotrons; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005583:Fourier Analysis; D006454:Hemoglobins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D039841:Leukocyte L1 Antigen Complex / Q000032:analysis / Q000378:metabolism; D008969:Molecular Sequence Data; D011392:Proline / Q000378:metabolism; D020033:Protein Isoforms / Q000032:analysis / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D011649:Pulmonary Alveolar Proteinosis / Q000378:metabolism*; D011650:Pulmonary Alveoli / Q000378:metabolism*; D037662:Pulmonary Surfactant-Associated Protein A / Q000032:analysis / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D006454:Hemoglobins; D039841:Leukocyte L1 Antigen Complex; D020033:Protein Isoforms; D020543:Proteome; D037662:Pulmonary Surfactant-Associated Protein A; D011392:Proline