title: [Impact of the palmitoylation of linker for activation of T cells on signal transduction pathway of CD59 in T cells].
abstract: OBJECTIVE
  To transfect Jurkat cells with linker for activation of T cells (LAT)-EGFP or LAT-EGFP-M (two cysteines of which near the juxtamembrane region were substituted for alanine) by electroporation, and observe the change in T cell signal transduction after the cross linkage with anti-CD59 mAb.
  
  
  METHODS
  By means of electroporation, Jurkat cells were transfected with LAT-EGFP or LAT-EGFP-M or PEGFP-N3 (control plasmid), and then subjected to cross linkage with anti-CD59 mAb. The locations of the fusion protein LAT-EGFP and LAT-EGFP-M were observed with confocal microscopy. The proliferation activity of the Jurkat cells was determined by MTT assay, and the content of interleukin 2 (IL-2) in cell supernatants was detected by ELISA. Western blotting was used to test the phosphorylation level of LAT in each group.
  
  
  RESULTS
  After the cross linkage of anti-CD59 mAb, LAT-EGFP-M could not locate on lipid rafts just as LAT-EGFP did. The cell proliferation and IL-2 secretion of Jurkat cells transfected with LAT-EGFP-M decreased as compared with the ones transfected with LAT-EGFP, EGFP-N3 or the untransfected ones. The phosphorylation level of LAT-EGFP-M was much lower than that of LAT-EGFP.
  
  
  CONCLUSIONS
  The mutation of palmitoylation site of LAT-EGFP attenuated the signal transduction of CD59 in T cells.
authors.affiliation: Department of Immunology, Medical College, Qingdao University, Qingdao 266071, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Qin
authors.forename: Meihua
authors.initials: M
authors.lastname: Gao
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
authors.forename: Bei
authors.initials: B
authors.lastname: Zhang
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Li
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Liang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D018957:CD59 Antigens / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D019169:Jurkat Cells; D054878:Lipoylation; D008565:Membrane Proteins / Q000378:metabolism*; D021381:Protein Transport; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D014162:Transfection
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018957:CD59 Antigens; D007376:Interleukin-2; C110810:LAT protein, human; D008565:Membrane Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins