関連する実験動画
Updated: Jun 20, 2026

09:16
Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
まとめ
タンパク質キナーゼCはppp60srcをセルリン12でin vivoでリン酸化する. 腫瘍プロモーターとダイアシルグリセロールによって誘発されるこの改変は,精製されたタンパク質キナーゼCを用いてインビトロで確認され,新しいリン酸化現象を強調した.
科学分野:
- 分子生物学は分子生物学である.
- セルラー・シグナリング
- 腫瘍学 腫瘍学
背景:
- pp60v-src (生肉肉腫ウイルス変異タンパク質) とpp60c-src (細胞同型) は,細胞成長における重要なタンパク質です.
- リン酸化は,タンパク質の機能の重要な規制メカニズムです.
研究 の 目的:
- 血清12でpp60srcのインビボリン酸化に責任を負うキナーゼを特定する.
- pp60src改変におけるタンパク質キナーゼCの役割を調査する.
主な方法:
- 腫瘍プロモーター (12-O-テトラデカノイルホルボル-13-アセテート,テレオシジン) とダイアシルグリセロールを用いたイン・ビボの酸化研究.
- 精製されたpp60c-src/pp60v-srcおよび様々なセリン/スレオニン特異のタンパク質キナーゼを用いたインビトロキナーゼアッセイ.
- 精製されたタンパク質キナーゼCを用いて pp60c-src N端を模倣する合成ペプチドのインビトロリン酸化.
主要な成果:
- 腫瘍プロモーターとダイアシルグリセロルは,ppp60srcをセルリン12で活体内で有意にリン酸化させた.
- 純化されたタンパク質キナーゼCのみがppp60c-srcとppp60v-srcをセルリン12でin vitroでリン酸化した.
- 精製されたタンパク質キナーゼCは,セリン12で合成ペプチドをリン酸化し,その活性部位を確認した.
結論:
- タンパク質キナーゼCは,ppp60srcをセルリン12でリン酸化させる主なキナーゼである.
- タンパク質キナーゼCによるこの新しいリン酸化イベントは,重要な生理学的影響を及ぼす可能性があります.
- セリン12のリン酸化による機能的影響を明らかにするために,さらなる研究が必要である.
関連する概念動画
Phosphorylation
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
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...
Covalently Linked Protein Regulators
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
These groups modify specific amino acids in a protein.
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...
PI3K/mTOR/AKT Signaling Pathway
The mammalian target of rapamycin (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast, mTORC2 consists of a rapamycin-insensitive companion...
cAMP-dependent Protein Kinase Pathways
Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...

