異なる受容体は,ATPとGTPの診断パターンを生み出します
Shawn C McCleskey1, Michael J Griffin, Stephen E Schneider
1University of Texas at Austin, Department of Chemistry and Biochemistry, Austin, Texas 78712, USA.
Journal of the American Chemical Society
|January 30, 2003
まとめ
この研究は,核酸塩酸塩を検出するための新しいセンサー配列を導入し,各分析物に対してユニークな指紋応答を提供します. この進歩により,アデノシン三リン酸および関連する化合物が含まれる溶液の選択的かつほぼリアルタイムなデジタル分析が可能になります.
科学分野:
- 化学センサー 化学センサー
- アナリティカル・ケミストリー (Analytical Chemistry) とは
- バイオテクノロジー バイオテクノロジー
背景:
- 核酸塩酸塩は,多数の細胞プロセスに関与する重要なバイオ分子です.
- 核酸塩酸塩の選択的かつ迅速な検出は,生化学的研究と診断に不可欠です.
- 既存の検出方法は,特定のアプリケーションに必要な感度,特異性,または速度を欠いている可能性があります.
研究 の 目的:
- ニュクレオチド・リン酸塩の選択的検出のための新しい結合配列センサーシステムを開発する.
- それぞれの分析物に対してユニークな"指紋"応答を提供できるセンサーシステムを構築する.
- 溶液中の核酸塩酸塩のほぼリアルタイムのデジタル分析を可能にします.
主な方法:
- 1,3,5-trisubstituted-2,4,6-triethylbenzene scaffoldとペプチドライブラリを使用した樹脂結合センサーのライブラリが合成されました.
- 化学センサーは,マイクロ機械化されたシリコンの空洞内の多成分センサー配列に統合されました.
- 光学的な変化は,デジタル分析のための充電結合装置を使用して監視されました.
- カロリメトリックシフトアッセイと時間依存型イメージング研究が行われました.
主要な成果:
- センサー配列は溶液中の核酸塩酸塩の選択性を示した.
- アデノシン5'-トリホスファート (ATP),アデノシン5'-モノホスファート (AMP),およびグアノシン5'-トリホスファート (GTP) を加えると異なる反応が観察されました.
- 主要コンポーネント分析は,ATP,GTP,AMPを区別するセンサーライブラリの能力を確認しました.
- 個々のセンサーは,因子負荷値に基づいて化学組成を明らかにするために配列化されました.
結論:
- 開発された組み合わせ配列センサーシステムは,選択的な核酸塩酸塩検出のための強力なツールを提供します.
- "指紋"応答は,分析物の堅固な差別化を可能にします.
- このアプローチは,ほぼリアルタイムのデジタル分析を容易にし,生化学的センサー能力を向上させます.
関連する概念動画
G-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.
GTPases and their Regulation
Guanine nucleotide-binding proteins (G-proteins), also known as GTPases, are a superfamily of proteins that regulate many cellular processes, such as cell signaling, vesicular transport, and the regulation of cell shape and motility. Mutation or dysfunction of these proteins can lead to disease. There are around 40,000 known G-proteins that can broadly be classified into two groups ‒ small G-proteins consisting of a single domain and large multi-domain G-proteins.
Large G-proteins, also known...
Large G-proteins, also known...
ATP Driven Pumps I: An Overview
ATP-driven pumps, also known as transport ATPases, are integral membrane proteins. They have binding sites for ATP located on the membrane's cytosolic side and the ion-conducting domain in the transmembrane region. These pumps use the free energy released from ATP hydrolysis to move the solutes across cell membranes against an electrochemical gradient.
There are four main types of ATP-driven pumps - P-type, V-type, F-type, and ABC transporter. All these pumps are of varying complexities and are...
There are four main types of ATP-driven pumps - P-type, V-type, F-type, and ABC transporter. All these pumps are of varying complexities and are...
ATP Driven Pumps II: P-type Pumps
The P-type pumps are a large family of integral membrane transporter ATPases. They are divided into five major types based on substrate specificity, from I to V.
A typical P-type pump has three cytosolic domains: nucleotide-binding (N), phosphorylation (P), and activator (A) domains. These domains are connected to the membrane-spanning helices by short amino acid segments. ATP hydrolysis and covalent phosphoenzyme intermediate formation are crucial parts of the catalytic cycle. At the highly...
A typical P-type pump has three cytosolic domains: nucleotide-binding (N), phosphorylation (P), and activator (A) domains. These domains are connected to the membrane-spanning helices by short amino acid segments. ATP hydrolysis and covalent phosphoenzyme intermediate formation are crucial parts of the catalytic cycle. At the highly...
Activation 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...
IP3/DAG Signaling Pathway
Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and produces two-second...


