Proteome Annotations and Identifications of the Human Pulmonary Fibroblast

Johan Malmström, Kristoffer Larsen, Lars Malmström, Ellen Tufvesson,§ Ken Parker, Jason Marchese, Brian Williamson, Steve Hattan, Dale Patterson, Steve Martin, Armin Graber, Peter Juhasz, Gunilla Westergren-Thorsson, and György Marko-Varga*
Cell & Molecular Biology, C13, BMC, 221 84 Lund, Sweden, Department of Electrical Measurement, Lund University, Box 124, 221 00 Lund, Sweden, Lung Division (Usil), Lund University, 221 85 Lund, Sweden, Applied Biosystems, 500 Old Connecticut Path, Framingham, Massachusetts 01701, and Analytical Chemistry, Lund University, Box 124, 221 00, Sweden
Journal of Proteome Research, 2004, 3 (3), pp 525–537
DOI: 10.1021/pr034104v
Publication Date (Web): March 26, 2004
Copyright © 2004 American Chemical Society

Abstract

Abstract Image

We hereby report on a three year project initiative undertaken by our research team encompassing large-scale protein expression profiling and annotations of human primary lung fibroblast cells. An overview is given of proteomic studies of the fibroblast target cell involved in several diseases such as asthma, idiopatic pulmonary disease, and COPD. It has been the objective within our research team to map and identify the protein expressions occurring in both activated-, as well as resting cell states. The JGGL database www.2DDB.org has been built around these data, allowing advanced hypothesis building using the interactive query bioinformatic tools developed. Gene ontology has been applied to these annotations, classifying and correlating protein expressions to function. The localization as well as the biological processes involved for the annotations are being presented including an annotation-, and sequence-identification strategy, resulting in close to 2000 protein identities. Both gel based, high resolution 2D-gels, and liquid-phase separation (three-dimensional HPLC), as well as the combination of gel- and LC-based approaches (1D-gels and nano-capillary LC, reversed-phase) were utilized. Protein sequencing and structure identities were acquired by a combination of MALDI-, and electrospray−mass spectrometry techniques. Phenotypical and morphological characterizations were also made for this human disease target cell in both stimulated- and resting-cell states. The use of functional assays that demonstrate the key regulating role of growth factors and cytokine stimuli such as PDGF, TGF-β, and EGF and the effect of ECM molecules such as Biglycan, are also presented and discussed.

Keywords: fibroblast • nucleus • proteome • lung • mass spectrometry • TGF-β

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  • Published In Issue June 14, 2004
  • Received November 10, 2003

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