相关实验视频
Updated: Aug 11, 2026

09:16
Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
概括
瘤促进剂12-O-tetradecanoyl-phorbol-13-acetate (TPA) 激活蛋白激酶C.TPA还诱导42K蛋白的氨酸酸化,这表明TPA在TPA中发挥了作用.
科学领域:
- 细胞生物学 细胞生物学
- 生物化学 生物化学
- 在瘤学瘤学.
背景情况:
- 12-O-tetradecanoyl-phorbol-13-acetate (TPA) 是一个强大的瘤促进体.
- 在体外,TPA激活蛋白激酶C (PKC),模仿二甲醇的作用.
- 在氨酸和氨酸残留物中,PKC化基质.
研究的目的:
- 研究TPA对培养纤维细胞的影响机制.
- 为了确定TPA是否通过氨酸酸化调解其作用.
- 为了检查一个特定的42K多的酸化状态,以对TPA做出反应.
主要方法:
- 用TPA处理培养的纤维细胞.
- 在氨酸残留物中对蛋白质酸化的分析.
- 研究外源添加的二甲基甘油的作用.
主要成果:
- 治疗TPA导致42,000分子量 (42K) 聚的氨酸酸化增加.
- 这种42K多也在响应EGF,PDGF和MSA等生长因子时被化为铁.
- 外源添加的二甲基甘油也刺激了这种42K蛋白的氨酸酸化.
结论:
- TPA的促进瘤效应可能包括诱导氨酸酸化.
- 42K聚是TPA,生长因子和二甲基甘油诱导的铁酸酸化的常见基质.
- 这一发现将TPA诱导的PKC激活与氨酸激酶信号通路联系起来.
相关概念视频
What are Second Messengers?
Because many receptor binding ligands are hydrophilic, they do not cross the cell membrane and thus their message must be relayed to a second messenger on the inside. There are several second messenger pathways, each with their own way of relaying information. G-protein coupled receptors can activate both phosphoinositol and cyclic AMP (cAMP) second messenger pathways. The phosphoinositol path is active when the receptor induces phospholipase C to hydrolyze the phospholipid,...
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...
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...
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...

