相关实验视频
Updated: Jul 13, 2026

11:13
Identification of Kinase-substrate Pairs Using High Throughput Screening
Published on: August 29, 2015
概括
研究pp60c-src中的氨酸酸化表明,Tyr 527中的低酸化和Tyr 416中的高酸化对于其充分的转化潜力至关重要.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- pp60c-src 是一种原瘤基因的氨酸激酶.
- 氨酸酸化在调节激酶活性和细胞转化方面发挥着至关重要的作用.
- 特定的氨酸残留物 (416,519,527) 是关键的监管区域.
研究的目的:
- 阐明在pp60c-src调节中特定的氨酸酸化位的作用.
- 确定Tyr 416,519和527中的修改如何影响pp60c-src活动和细胞转化.
主要方法:
- 用于取代ppp60c-src的416位,519位和527位的氨酸替代氨酸,使用了局部导向的突变发生.
- 进行了对形态转变,软生长和焦点形成的测试.
- 在实验室中对突变pp60c-src蛋白进行了激酶活性测定.
主要成果:
- 具有Tyr 527 (单独或与Tyr 519) 修改的突变者表现出形态转变,软生长和焦点形成.
- 这些突变还显示出高的体外激酶活性.
- 在Tyr 416的替代部分抑制了激酶活性,但在Tyr 527修改的背景下取消了转变表型.
结论:
- pp60c-src 的全部转化潜力需要在 Tyr 527 中进行低酸化.
- 在Tyr 416中,高酸化对于诱导焦点形成和软成长至关重要.
- Tyr 527和Tyr 416的酸化状态都对pp60c-src介导的细胞转化至关重要.
相关概念视频
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...
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.
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...
Receptor Tyrosine Kinases
Receptor tyrosine kinases or RTKs are membrane-bound receptors that phosphorylate specific tyrosine on protein substrates. RTKs regulate cellular growth, differentiation, survival, and migration. They contain an extracellular ligand binding domain, a transmembrane domain, and a cytosolic tail with intrinsic kinase activity. Several extracellular signaling molecules activate RTKs in one or more ways and relay the signal downstream. Ligands such as platelet-derived growth factor (PDGF) or...

