相关实验视频
Updated: Jul 10, 2026

14:57
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
概括
将突变引入的pp60c-src揭示了改变氨酸527会增加激酶活性和转化潜力. 炭基末端对于与多瘤中瘤抗原 (mT) 的关联至关重要.
科学领域:
- 分子生物学分子生物学
- 瘤发生的发生因子.
- 蛋白激酶的功能 蛋白激酶的功能
背景情况:
- 肉pp60c-src是一种由其碳氧终端区域调节的原致瘤氨酸激酶.
- 在氨酸527 (Y527) 的酸化通常会抑制激酶活性.
- 影响这种调节区域的突变可以改变SRC激酶功能和细胞转化.
研究的目的:
- 为了研究 pp60c-src.src.的碳氧末端调节区域的特定突变的功能后果.
- 确定这些突变对蛋白激酶活性,体内酸化和细胞转化的影响.
- 探索卡基末端在与多瘤中瘤抗原 (mT) 相互作用中的作用.
主要方法:
- 用局部定向突变发生来引入两种突变进入的pp60c-src基因:F527 (Y527F) 和Am517 (切断).
- 突变型和野生型蛋白质在活体中被分析到氨酸416 (Y416) 的酸化.
- 测量了蛋白激酶的特定活性,并对NIH 3T3细胞进行了焦点形成测定.
- 评估了pp60c-src变体和多瘤中瘤抗原 (mT) 之间的复杂形成.
主要成果:
- 在体内,F527和Am517突变pp60c-src蛋白都在Y416酸化.
- 与野生类型相比,F527突变体的特定激酶活性增加了5到10倍.
- Am517突变体表现出与野生类型相似的激酶活性,但这两种突变体都诱导了焦点形成,频率较低,焦点较mt小.
- 野生型和F527 pp60c-src与mt形成复合物,而Am517 pp60c-src没有.
结论:
- 不能酸化Y527显著增强pp60c-src蛋白激酶活性和转化能力.
- 由mT诱导的转化涉及到细胞事件,除了在pp60c-src上缺乏Y527酸化之外.
- 激活pp60c-src激酶活性可能不必由Am517突变介导的转化.
- pp60c-src 的碳素末端是必要的,以使其与mT相关联.
相关概念视频
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.
Abnormal Proliferation
Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...
Cancer-Critical Genes I: Proto-oncogenes
Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
The Ras Gene
The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a superfamily...
Ras is a superfamily...
mTOR Signaling and Cancer Progression
The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
The mTOR pathway or the...
Cancer-Critical Genes I: Proto-oncogenes
Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...

