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Mapping and identification of interferon gamma-regulated HeLa cell proteins separated by immobilized pH gradient two-dimensional gel electrophoresis.
Shaw, A C; Røssel Larsen, M; Roepstorff, P; Justesen, J; Christiansen, G; Birkelund, S.
Afiliación
  • Shaw AC; Department of Medical Microbiology and Immunology, University of Aarhus, Denmark. shaw@medmicro.aau.dk
Electrophoresis ; 20(4-5): 984-93, 1999.
Article en En | MEDLINE | ID: mdl-10344276
Interferon gamma (IFN-gamma) is a potent immunomodulatory lymphokine, secreted by activated T-lymphocytes and NK-cells during the cellular immune response. Actions of IFN-gamma are mediated through binding to the IFN-gamma-receptor, present on most cells, and the subsequent activation of a great magnitude of IFN-gamma responsive genes has been reported previously. Our goal is to identify and map IFN-gamma-regulated HeLa cell proteins to the two-dimensional polyacrylamide gel electrophoresis with the immobilized pH gradient (IPG) two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) system. A semiconfluent layer of HeLa cells was grown on tissue culture plates, and changes in protein expression due to 100 U/mL IFN-gamma were investigated at different periods after treatment, using pulse labeling with [35S]methionine/cysteine in combination with 2-D PAGE (IPG). The identity of eight protein spots was elucidated by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS), and several variants of the IFN-gamma-inducible tryptophanyl-tRNA synthetase (hWRS) were detected by immunoblotting.
Asunto(s)
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Mapeo Peptídico / Cisteína Endopeptidasas / Electroforesis en Gel Bidimensional / Proteínas / Interferón gamma / Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Electrophoresis Año: 1999 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Alemania
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Mapeo Peptídico / Cisteína Endopeptidasas / Electroforesis en Gel Bidimensional / Proteínas / Interferón gamma / Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Electrophoresis Año: 1999 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Alemania