Transmembrane TNF (pro‐TNF) is palmitoylated

FEBS Letters

Takami, Kenji¹  Tanaka, Kenji¹  Utsumi, Toshihiko¹  Ishisaka, Rumi¹  Kira, Yukimi¹  Klostergaard, Jim²  Takeshige, Tomonori¹ 

[1] Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan;Department of Molecular and Cellular Oncology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030, USA

关键词: Tumor necrosis factor;    Transmembrane tumor necrosis factor;    Protein palmitoylation;    Protein acylation;    Post-translational modification;    Signal transduction;    TNF;    tumor necrosis factor;    PCR;    polymerase chain reaction;    ECL;    enhanced chemiluminescence;    DPBS;    Dulbecco's phosphate-buffered saline;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    LAT;    linker for activation of T-cells;   

DOI  :  10.1016/S0014-5793(01)02576-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Human transmembrane tumor necrosis factor (pro-TNF) was examined for protein acylation. The cDNA encoding pro-TNF was expressed in both COS-1 cells and Sf9 cells and metabolic labeling with [³H]myristic or [³H]palmitic acid was attempted. The 17 kDa mature TNF secreted from the transfected cells was not labeled, whereas the 26 kDa pro-TNF was specifically labeled with [³H]palmitic acid. The [³H]palmitic acid labeling of pro-TNF was eliminated by treatment with hydroxylamine, indicating that the labeling was due to palmitoylation of a cysteine residue via a thioester bond. Site-directed mutagenesis of the two cysteine residues residing in the leader sequence of pro-TNF demonstrated that palmitoylation of pro-TNF occurs solely at Cys-47, located at the boundary between the transmembrane and cytoplasmic domains of pro-TNF. Thus, pro-TNF interacts with the plasma membrane via both its proteinaceous transmembrane domain and a lipid anchor.

【 授权许可】

Unknown