分子クローニングとヒト55 kd腫瘍死滅因子受容体の発現
H Loetscher1, Y C Pan, H W Lahm
1Central Research Units, F. Hoffmann-LaRoche LTD, Basel, Switzerland.
Cell
|April 20, 1990
まとめ
研究者らは55kdの腫瘍死滅因子 (TNF) 受容体を浄化し,そのアミノ酸配列を決定した. この研究は,TNF受容体を特定しました.
科学分野:
- 分子生物学は分子生物学である.
- 免疫学 免疫学とは
- 細胞生物学 細胞生物学
背景:
- 腫瘍死滅因子 (TNF) の2つの異なる受容体55kdと75kdは,様々な細胞で低いレベルで発現しています.
- TNF受容体の構造と機能を理解することは,炎症と免疫反応を理解するために不可欠です.
研究 の 目的:
- 55kdのTNF受容体を浄化し特徴づけること.
- TNF受容体の機能の分子基礎を決定し,潜在的な治療標的を特定する.
主な方法:
- HL60細胞から55kdのTNF受容体の浄化.
- 部分アミノ酸配列の決定.
- 変性オリゴヌクレオチドプライマーを使用してcDNA断片を放大する逆転写酵素-ポリメラーゼ連鎖反応 (RT-PCR).
- 増幅された断片をプローブとして使用した人間の胎盤のcDNAライブラリのスクリーニング.
- 複製されたcDNAをCOS-1細胞とバキュロウイルスシステムで発現させ,TNF結合特性を評価する.
主要な成果:
- 55kdのTNF受容体の部分アミノ酸配列が決定されました.
- 66bpのcDNA断片が増幅され,人間の胎盤のcDNAライブラリから重複するクローンを識別するために使用されました.
- 全長cDNAは455アミノ酸のTNF受容体タンパク質を予測し,異なる機能ドメイン (リーダー,細胞外,膜外,細胞内) を有する.
- 発現したTNF受容体は,ネイティブ受容体と同等のTNF結合特性を授与した.
- 55kd TNF受容体とNGF受容体の細胞外ドメインの間に有意な配列ホモロジーが観察されました.
結論:
- この研究では,55kdのTNF受容体を成功裏に浄化し,特徴づけ,その分子構造を明らかにしました.
- この発見は,TNF受容体のシグナル伝達経路と疾患におけるその役割を理解するための基礎を提供します.
- NGF受容体との同質性は,潜在的に共有されたシグナル伝達機構または進化的関係を示唆する.
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Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
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Loss of Tumor Suppressor Gene Functions
Tumor suppressor genes are normal genes that can slow down cell division, repair DNA mistakes, or program the cells for apoptosis in case of irreparable damage. Hence, they play an essential role in preventing the proliferation of damaged cells.
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