通过结合诱导的DNA条形码转移标记,精确地绘制直接膜蛋白相互作用的精确地图
Weikang Li1, Xigao Chen2, Nachuan Wen1,3
1Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan 410082, China.
Nano letters
|August 4, 2025
概括
这项研究引入了一种新的方法来追踪直接蛋白质相互作用,如PD1/PD-L1结合,通过细胞之间转移DNA条形码. 这允许精确监测细胞-细胞通信及其对T细胞重编程的影响.
科学领域:
- 分子生物学分子生物学
- 免疫学 免疫学 免疫学
- 生物化学 生物化学
背景情况:
- 细胞间的通信依赖于动态的膜蛋白相互作用.
- 准确跟踪直接蛋白质结合对于理解生物过程至关重要.
- 现有的近距离标记方法难以区分直接结合与非特异性相互作用.
研究的目的:
- 开发一种结合诱导的标签策略,用于在相互作用的蛋白质之间精确的1:1DNA条形码传输.
- 为了使单个蛋白质-蛋白质结合事件在细胞界面上的敏感和顺序跟踪.
- 研究代蛋白结合对细胞重编程的功能结果.
主要方法:
- 设计了功能化的阿普坦体探针,针对癌细胞上的PD-L1.
- 利用点击化学,在结合后在T细胞上从PD-L1转移到PD1的共价DNA.
- 集成的条形码原始序列和DNA信号放大用于敏感检测.
- 结合单细胞转录分析,分析T细胞重编程.
主要成果:
- 成功展示了一种绑定诱导的标签策略,用于跟踪蛋白质相互作用.
- 实现了个别PD1/PD-L1结合事件的敏感和顺序监测.
- 通过综合单细胞分析揭示了代PD1/PD-L1结合对T细胞重编程的渐进性影响.
结论:
- 开发的平台可以精确跟踪细胞界面的直接蛋白质-蛋白质相互作用.
- 这种方法为解码复杂的细胞-细胞通信动态提供了强大的工具.
- 这些发现提供了关于免疫检查点相互作用对T细胞行为的功能后果的见解.
相关概念视频
02:57Southern Blot
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
02:34DNA Microarrays
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...