相关实验视频

Updated: Jun 2, 2025

Assaying Protein Kinase Activity with Radiolabeled ATP

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

在ERK激酶转位报告器上的CDK2活动交叉声可以通过计算解决

Timothy E Hoffman¹, Chengzhe Tian¹, Varuna Nangia¹

¹Department of Biochemistry & BioFrontiers Institute, University of Colorado, Boulder, CO 80303, USA.

|January 16, 2025

概括

研究人员发现,ERK激酶转位报告器 (KTR) 可以不准确地反映细胞循环进展,这是由于循环素依赖激酶2 (CDK2) 活性. 开发了计算方法来纠正这种交叉对准MAPK信号分析的交叉.

科学领域:

蜂信号传输是如何进行的
分子生物学分子生物学
生物化学生物化学

背景情况:

线素激活蛋白激酶 (MAPK) 途径,包括细胞外信号调节激酶 (ERK),调节细胞增殖.
酶转位报告器 (KTR) 通常用于测量活细胞中的ERK活性.

研究的目的:

调查影响ERK活动报告员的潜在交叉通话.
开发精确的方法来量化循环细胞中的MAPK活性.

主要方法:

单细胞时隔成像技术.
使用了选择性循环素依赖性激酶2 (CDK2) 抑制剂.
开发了计算校正方法 (线性和非线性).
将ERK KTR与基于FRET的ERK传感器 (EKAREN5) 进行比较.

主要成果:

ERK KTR部分感知CDK2活动,导致细胞周期进展期间明显的ERK活动增加.
CDK2交叉被抑制剂研究和EKAREN5传感器中缺乏交叉证实.
一个相关的p38 KTR也显示了部分的CDK2活动影响.
计算校正方法成功地从KTRs中减去了CDK2信号.

结论:

ERK和p38 KTRs与CDK2呈现交叉声,影响循环细胞中的测量.
开发的计算工具可以更准确地量化MAPK活动.
这些发现对于精确分析活跃增殖细胞中MAPK信号的关键.

关键词:

这是一种CDK2抑制剂.这是一个CDK2传感器.EKAREN5 FRET的记者是什么意思埃尔克 KTRTR 的时间这是一种MAPK抑制剂.酶转位报告员的转位报告员传感器的交叉声道传感器的交叉声道一个单细胞追踪系统.时间间隔显微镜技术

更多相关视频

Spatial and Temporal Analysis of Active ERK in the C. elegans Germline

Spatial and Temporal Analysis of Active ERK in the C. elegans Germline

Published on: November 29, 2016

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Published on: January 27, 2012

相关概念视频

M-Cdk Drives Transition Into Mitosis

M-Cdk Drives Transition Into Mitosis

Checkpoints throughout the cell cycle serve as safeguards and gatekeepers, allowing the cell cycle to progress in favorable conditions and slow or halt it in problematic ones. This regulation is known as the cell cycle control system.
Cyclin-dependent kinases, or Cdks, work in concert with cyclins to control cell cycle transitions. M-Cdk, a complex of Cdk1 bound to M cyclin, is a well-known example of this coordinated control that drives the transition from the G2 to the M phase.
M cyclin...

Inhibition of Cdk Activity

Inhibition of Cdk Activity

The orderly progression of the cell cycle depends on the activation of Cdk protein by binding to its cyclin partner. However, the cell cycle must be restricted when undergoing abnormal changes. Most cancers correlate to the deregulated cell cycle, and since Cdks are a central component of the cell cycle, Cdk inhibitors are extensively studied to develop anticancer agents. For instance, cyclin D associates with several Cdks, such as Cdk 4/6, to form an active complex. The cyclin D-Cdk4/6 complex...

Positive Regulator Molecules

Positive Regulator Molecules

Mitotic cell division results in daughter cells that exactly resemble the parent cell. However, errors in the DNA replication or distribution of genetic material may lead to genetic mutations that may be passed down to every new cell formed from the resulting abnormal cell. Propagation of such mutant cells is restricted through checkpoint mechanisms present at different stages of the cell cycle. These checkpoints involve regulator molecules that either promote or demote cell cycle events.

MAPK Signaling Cascades

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...

S-Cdk Initiates DNA Replication

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of...

DNA Damage can Stall the Cell Cycle

DNA Damage can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...