一个基因编码的红外探测器
Kathryn C Schultz1, Lubica Supekova, Youngha Ryu
1Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. schultz@scripps.edu
Journal of the American Chemical Society
|October 26, 2006
概括
研究人员开发了一种方法,利用大肠杆菌将一种新型氨基酸 - - 氨基氨酸 (pCNPhe) 纳入蛋白质中. 该技术允许对蛋白质结构和相互作用进行敏感的红外监测.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 生物物理学的生物物理.
背景情况:
- 将非正规氨基酸 (ncAA) 具体地纳入蛋白质对于扩大蛋白质功能和启用新型分析技术至关重要.
- 现有的ncAA整合方法经常面临效率和特异性方面的挑战.
- 珀色无意义编码子 (TAG) 是遗传码扩展的常见目标.
研究的目的:
- 进化一个正交tRNA/aminoacyl-tRNA合成酶对,以有效和选择性地将para-cyanophenylalanine (pCNPhe) 纳入大肠杆菌中的蛋白质.
- 使用pCNPhe作为振动探针来研究蛋白质结构和动态.
主要方法:
- 对pCNPhe特有的正交tRNA/aminoacyl-tRNA合成酶对的演变.
- 在大肠杆菌中,pCNPhe在珀色 (TAG) 位点的蛋白质中被遗传纳入.
- 对PCN的光谱分析he-替代的肌球蛋白用于连接体结合的研究.
主要成果:
- 成功进化了一个正交tRNA/aminoacyl-tRNA合成酶对,使有效的pCNPhe结合成为可能.
- 在肌球蛋白中证明了pCNPhe对histidine-64的特定位点替代.
- 展示了pCNPhe作为灵敏的红外线报道器,用于带结合事件.
结论:
- 开发的方法提供了一个强大的系统,用于将pCNPhe具体纳入蛋白质.
- pCNPhe作为一种有价值的红外探测器,用于研究蛋白质结构,生物分子相互作用和结构变化.
相关概念视频
03:31Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...