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Updated: Jul 29, 2026
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Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
Published on: June 9, 2017
Gタンパク質結合受容体であるヒトバソプレシンV2受容体の溶液NMRスペクトロスコーピー
Changlin Tian1, Richard M Breyer, Hak Jun Kim
1Center for Structural Biology and Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-8725, USA.
Journal of the American Chemical Society
|June 2, 2005
まとめ
重要なGタンパク質結合受容体 (GPCR) であるヒトバソプレシンV2受容体について,高品質のNMRスペクトルが得られた. この画期的な発見により,GPCRの詳細な構造研究が可能になり,健康と病気を理解する上で極めて重要です.
科学分野:
- バイオケミストリー バイオケミストリー
- 構造生物学 構造生物学とは
- 薬理学 薬理学とは
背景:
- Gタンパク質結合受容体 (GPCR) は,多くの生理学的プロセスに関与する重要な膜タンパク質です.
- GPCRの機能障害は,様々なヒト疾患に関与しており,重要な薬物標的となっています.
- GPCRの構造を理解することは,合理的な薬剤設計と治療開発に不可欠です.
研究 の 目的:
- 人間のGPCR,すなわちバソプレシンV2受容体に対する核磁気共鳴 (NMR) スペクトルの解像度を示します.
- 受容体の共鳴の過半数のバックボーン共鳴の割り当ての実現可能性を実証する.
- 膜に結合したGPCRから高品質のNMRデータを取得するための堅牢な方法の確立.
主な方法:
- 膜内のバソプレシンV2受容体の発現,インクルージョンボディ形成を回避する.
- 生物化学的に軽度のリソリピド洗剤を使用して受容体の抽出と浄化.
- 溶液NMRスペクトロスコーピーのための洗剤ミセルでNMRサンプルを準備する.
主要な成果:
- ヒトのバソプレシンV2受容体の溶液NMRスペクトルの良好な解像度を達成しました.
- 脊髄共鳴の割り当ては,ほとんどの共鳴で実現可能であることを実証しました.
- 高品質のスペクトルに不可欠な表現,抽出,および洗浄剤の特定の組み合わせを特定しました.
結論:
- 提示された方法論は,NMRを用いたGPCRの高解像度構造研究を可能にします.
- この研究は,GPCRの構造と機能の関係に関する将来の調査のための基礎を提供します.
- この発見は,GPCRを標的とした薬剤の発見にとって重要である.
関連する概念動画
01:21G-protein Coupled Receptors
G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
01:15G Protein-coupled Receptors
G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
01:22Activation and Inactivation of G Proteins
Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high affinity and are together...
01:12GPCR Desensitization
G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...
01:30Transducer Mechanism: G Protein–Coupled Receptors
G Protein–Coupled Receptors (GPCRs) are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to various stimuli. GPCRs regulate critical physiological pathways and are excellent drug targets for treating diseases such as diabetes, cancer, obesity, depression, or Alzheimer's. Nearly 35% of approved drugs implement their therapeutic effects by selectively interacting with specific GPCRs.
GPCRs are also called heptahelical, 7TM, or...
GPCRs are also called heptahelical, 7TM, or...
01:27Transducer Mechanism: Enzyme-Linked Receptors
Enzyme-linked receptors are cell-surface receptors acting as an enzyme or associating with an enzyme intracellularly. They make excellent drug targets. Drugs can bind to the extracellular ligand-binding domain or directly affect their enzymatic domain and alter their activity.
Major types that are helpful drug targets include:
Major types that are helpful drug targets include: