Electrophoresis · 2001 · 54 citations · 22 references
Life SciencesCell BiologyImmunologyGenomicsProtein BiochemistryProteomicsBottom-up ProteomicsMass Spectrometry Data AnalysisMass Spectrometry-based ProteomicsProtein Database SearchCd4 Positive T CellsCd4 T Cell ActivationPeptide-mass FingerprintingProteome AnnotationT Helper CellsTime-of-flight Mass SpectrometryTwo-dimensional ElectrophoresisTwo-dimensional Gel Electrophoresis
We have established the first public database of human primary T helper cell proteome using two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight-mass spectrometry. For the database, CD4+ human T cells were activated with anti-CD3+anti-CD28 antibodies and metabolically labeled with [35S]methionine for 24 h. Cells were lysed and proteins were separated by 2-DE. About 1500 protein spots were detected in the resulting 2-DE gels with silver staining, and 2000 spots with autoradiography. We have identified 91 proteins from the 2-DE gels using peptide mass fingerprinting, and annotated them to our database. The identified proteins are also linked to SWISS-PROTand NCBI protein databases. Our database is available via the Internet at http://www3.btk.utu.fi:8080/Genomics/Proteomics/Database.
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