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Proteome annotations and identifications of the human pulmonary fibroblast.

Type Information
Nr 6 (Research article)
Authors Malmström, Johan; Larsen, Kristoffer; Malmström, Lars; Tufvesson, Ellen; Parker, Ken; Marchese, Jason; Williamson, Brian; Hattan, Steve; Patterson, Dale; Martin, Steve; Graber, Armin; Juhasz, H; Westergren-Thorsson, Gunilla; Marko-Varga, György
Title Proteome annotations and identifications of the human pulmonary fibroblast.
Journal J Proteome Res (2004) 3 525-37
DOI 10.1021/pr034104v
Citations 41 citations (journal impact: 5.68)
Abstract 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-beta and EGF and the effect of ECM molecules such as Biglycan are also presented and discussed.