Mapping and identification of interferon gamma‐regulated HeLa cell proteins separated by immobilized pH gradient two‐dimensional gel electrophoresis

Interferon gamma (IFN‐γ) is a potent immunomodulatory lymphokine, secreted by activated T‐lymphocytes and NK‐cells during the cellular immune response. Actions of IFN‐γ are mediated through binding to the IFN‐γ‐receptor, present on most cells, and the subsequent activation of a great magnitude of IFN‐γ responsive genes has been reported previously. Our goal is to identify and map IFN‐γ‐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‐γ 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‐γ‐inducible tryptophanyl‐tRNA synthetase (hWRS) were detected by immunoblotting.

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