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Updated: Jul 6, 2026

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Identification of Kinase-substrate Pairs Using High Throughput Screening
Published on: August 29, 2015
信号伝達ネットワークにおけるグローバル,インビヴォ,およびサイト固有のリン酸化ダイナミクス
Jesper V Olsen1, Blagoy Blagoev, Florian Gnad
1Center for Experimental BioInformatics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense, Denmark.
Cell
|November 4, 2006
まとめ
この研究では,タンパク質のリン酸化部位をマッピングするための質量スペクトロメトリー法が導入されています. これは,表皮成長因子 (EGF) の刺激がこれらの部位をダイナミックに変化させ,細胞シグナル伝達ネットワークの洞察を提供する方法を示しています.
科学分野:
- 細胞生物学 細胞生物学
- バイオケミストリー バイオケミストリー
- プロテオミクス プロテオミクスは,プロテオミクスの
背景:
- 細胞シグナル伝達は,翻訳後のタンパク質改変,主に可逆性タンパク質リン酸化に依存しています.
- ダイナミックなリン酸化変化を理解することは,細胞の調節を解読する上で極めて重要です.
研究 の 目的:
- リン酸化部位を特定し,定量化するための質量スペクトロメトリー技術を開発し,適用する.
- エピデルマ・成長因子 (EGF) 刺激への反応として,リン酸化の時間的動態を分析する.
- ダイナミックなフォスフォプロテオームデータに関する包括的なデータベース (Phosida) を作成する.
主な方法:
- リン酸化部位の特定と定量化のために質量スペクトロメトリを活用した.
- 動的変化を観察するために,上皮成長因子 (EGF) を含むHeLa細胞を刺激した.
- リン酸化部位とその細胞下部位の時間動態を記録し,分析した.
主要な成果:
- 2,244のタンパク質に 6,600のリン酸化部位を特定した.
- フォシダデータベースのEGF刺激後のこれらの場所のタイムダイナミクスを文書化しています.
- 場所の14%がEGFによる少なくとも2倍の調節を示しており,時間的なプロファイルが異なっています.
- ほとんどのタンパク質には,異なる動力学を持つ複数のリン酸化部位があり,信号統合プラットフォームを示唆していることが観察されました.
- 特定された標的には,キナーゼ,ウビキチンリガゼ,グアニンヌクレオチド交換因子,および転写レギュレータが含まれています.
結論:
- ダイナミックなフォスフォプロテオームは,細胞の調節に関する全局的な見解のための重要なリンクを提供します.
- タンパク質の複数のリン酸化部位は,細胞信号を統合するためのプラットフォームとして機能します.
- この技術は,刺激に対する反応として,リン酸化動態の包括的な分析を可能にします.
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関連する概念動画
Protein Kinases and Phosphatases
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein Kinases and Phosphatases
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Assembly of Signaling Complexes
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Interactions Between Signaling Pathways
Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
Amplifying Signals via Second Messengers
Many receptor binding ligands are hydrophilic; they do not cross the cell membrane but bind to cell-surface receptors. Thus, their message must be relayed by second messengers present in the cell cytoplasm. There are several second messenger pathways, each with its own way of relaying information. For example, the G protein-coupled receptors can activate both phosphoinositol and cyclic AMP (cAMP) second messenger pathways. The phosphoinositol pathway is active when the receptor induces...
Amplifying Signals via Enzymatic Cascade
When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...

