相关实验视频
Updated: Jun 20, 2026

09:16
Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
概括
在体内,蛋白质激酶C在血清12的pp60src中酸化. 这种由瘤促进剂和二甲糖醇诱导的修饰在体外使用纯化蛋白激酶C证实,突出显示了一种新的酸化事件.
科学领域:
- 分子生物学分子生物学
- 细胞信号传输 细胞信号传输
- 在瘤学瘤学.
背景情况:
- pp60v-src (肉瘤病毒转化蛋白) 和pp60c-src (细胞同类物) 是细胞生长中的关键蛋白质.
- 酸化是蛋白质功能的关键调节机制.
研究的目的:
- 确定负责pp60src在血清12中的体内酸化的激酶.
- 为了研究蛋白质激酶C在pp60src修饰中的作用.
主要方法:
- 在体内化研究使用瘤促进剂 (12-O-tetradecanoylphorbol-13-乙酸盐,teleocidin) 和dyacylglycerol.
- 在体外激酶测试用纯化的pp60c-src/pp60v-src和各种氨酸/氨酸特异性蛋白激酶.
- 在试验室中化模仿pp60c-src N端的合成,使用纯化的蛋白激酶C.
主要成果:
- 瘤促进剂和糖醇在体内诱导了显著的pp60src酸化在血清12.
- 只有纯化蛋白激酶C在体内在12级血清中化pp60c-src和pp60v-src.
- 纯化蛋白激酶C在12中酸化了一种合成,证实了其活性部位.
结论:
- 蛋白质激酶C是负责pp60src酸化的主要激酶,在生物体内在12级血清中酸化.
- 这种新型的酸化事件由蛋白激酶C可能具有显著的生理影响.
- 需要进一步的研究来阐明12酸化的功能后果.
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...
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,...

