Abstract

European Journal of Mass Spectrometry
Volume 14 Issue 6, Pages 379–390 (2008)
doi: 10.1255/ejms.957

Aberrant processing forms of lung surfactant proteins SP–B and SP–C revealed by high-resolution mass spectrometry

Dmitry Galetskiy,^a Markus Woischnik,^b Jan Ripper,^b Matthias Griese^b and Michael Przybylski^a,*
aLaboratory of Analytical Chemistry and Biopolymer Structure Analysis, Department of Chemistry, University of Konstanz, Box M 73178457 Konstanz, Germany. E-mail: Michael.Przybylski@uni- konstanz.de
^bDepartment of Pediatric Pneumology, Dr. von Haunersches Kinderspital, University of Muenchen, Lindwurmstr. 4, 80337 Muenchen, Germany

The mutation (g.1286T>C) of the pulmonary surfactant-associated protein C gene (SFTPC) leads to the I73T substitution in the precursor protein (pro–SP–C) and results in interstitial lung disease with the histological pattern of non-specific interstitial pneumonia and pulmonary alveolar proteinosis. Central for the disease is the abnormal processing of the SP–C pro–protein to mature SP–C; however little is known about the nature of intermediates and processing products. We report here the application of high resolution Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry to the characterization of processing intermediates of hydrophobic pulmonary surfactant proteins SP–B and SP–C in intra-alveolar surfactant material of a patient with I73T mutation. SP–C and SP–B processing forms were separated from broncho-alveolar lavage fluid using chloroform/methanol extraction and sodium dodecyl sulfate poly acrylamide gel electrophoreis, detected by Western blot and identified by electrospray- and matrix-assisted laser desorption/ionization- FT-ICR mass spectrometry. The mass spectrometric and immuno-analytical results show the intra-alveolar accumulation of an aberrant C-terminal SP–C processing products in which the mature SP–C protein part is missing and aberrant processing intermediates of SP–B.

Keywords: pulmonary alveolar proteinosis, lung surfactant proteins, bronchoalveolar lavage fluid, FT-ICR mass spectrometry, proteome analysis, SP–C, SP–B precursor proteins, mutation forms

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