Analyzing mitogen-activated protein kinase (MAPK) activities in T cells.
The recognition of peptide antigens by T cells through their antigen receptors (T cell receptors, or TCR), together with the ligation of additional surface molecules called costimulatory receptors, rapidly induces interactive signaling pathways that lead to transcriptional initiation at genes such as that of the autocrine growth factor interleukin 2 (IL-2). Activation of the ERK and JNK subfamilies of MAPK mediates some of these signals. This unit presents procedures for a solid-phase kinase assay and immune-complex kinase assay to measure JNK and ERK activities, respectively, in T cells that have been appropriately stimulated. Also described is a procedure for preparing GSThyphen;cJun/GSH-Sepharose beads needed in the solid-phase JNK protein kinase activity assay.
Whitmarsh
Alan J
AJ
University of Manchester, School of Biological Sciences, Manchester, United Kingdom.
Davis
Roger J
RJ
D000818:Animals; D019898:Autocrine Communication / Q000502:physiology; D007118:Immunoassay; D007376:Interleukin-2 / Q000276:immunology / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000276:immunology / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000201:enzymology* / Q000276:immunology; D014158:Transcription, Genetic / Q000502:physiology
D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D020928:Mitogen-Activated Protein Kinases