| Project/Area Number |
12660080
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Yamaguchi University |
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Principal Investigator |
UTSUMI Toshihiko Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (20168727)
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| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | tumor necrosis factor (TNF) / processing / posttranslational modification |
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| Research Abstract |
Human transmembrane tumor necrosis factor (pro-TNF) was examined for posttranslational protein modification. The cDNA coding for 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 thiocster 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.
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