爱皮托普:预测和验证非破坏性的爱皮托普标记站点.
Joseph Zinski1, Parnal Joshi2,3, Henri Chung2,3
1Department of Cell and Development Biology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
概括
EpicTope识别了用于表位标记的最佳蛋白质位点,确保蛋白质功能得到维持. 这种工具有助于蛋白质的体内鉴定,跟踪和净化.
科学领域:
- 分子生物学分子生物学
- 生物化学 生化学
- 遗传学 是一个遗传学.
背景情况:
- 在体内蛋白质分析中,尾标记至关重要.
- 识别最佳的标签插入位是具有挑战性的,并影响蛋白质功能.
研究的目的:
- 开发EpicTope,这是一个计算工具,用于预测理想的表位标签插入位置.
- 为了尽量减少在表位标记期间对蛋白质功能的干扰.
主要方法:
- 埃皮克托普采用评分功能,考虑二级/三级结构,溶剂可访问性和无序区域.
- 在斑马鱼Smad5和Hdac1蛋白质上得到验证.
主要成果:
- 内部标记的Smad5和Hdac1蛋白质拯救了各自的突变斑马鱼胚胎.
- N-和C-终端标签表现出不同的成功,与EpicTope预测保持一致.
- 标记的蛋白质可以通过免疫光,西式涂抹和免疫沉来检测抗体.
结论:
- EpicTope有效地识别了功能性表皮质标签插入部位.
- 该工具有助于成功的体内蛋白质分析和净化.
相关概念视频
Tagging and Fusion Proteins
8.7K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
8.7K
Conserved Binding Sites
5.3K
Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
5.3K


