对于分裂蛋白质蛋白酶传感器的一种自抑制的卷绕卷绕设计策略
Sujan S Shekhawat1, Jason R Porter, Akshay Sriprasad
1Department of Chemistry & Biochemistry, University of Arizona, Tucson, Arizona 85721, USA.
Journal of the American Chemical Society
|October 7, 2009
概括
研究人员开发了新的遗传编码生物传感器,这些生物传感器在蛋白酶活动时被激活. 这种新的自抑制设计为检测像caspase-3这样的蛋白酶提供了1000倍的信号增强.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 蛋白酶是细胞过程的关键调节者,包括细胞循环控制和细胞亡.
- 开发精确的工具来监测蛋白酶活性对于生物研究和治疗开发至关重要.
研究的目的:
- 设计和优化一种新的基因编码,开启蛋白酶生物传感器家族.
- 为了建立一个一般的自抑制线圈-线圈策略来控制碎片化的蛋白质活性.
主要方法:
- 设计了一种自抑制的线圈-线圈开关,该开关在蛋白质分解裂变时激活.
- 使用分裂蛋白记者 (火虫光酶和β-乳糖酶) 来产生信号.
- 优化了三代生物传感器,并使用TEV蛋白酶和caspase-3进行了测试.
主要成果:
- 在TEV蛋白酶添加后,通过最终一代生物传感器实现了生物发光信号的1000倍增加.
- 证明了这种方法的多功能性,使用不同的分裂蛋白记者和治疗相关的蛋白酶 (caspase-3).
- 开发了一种双蛋白酶传感器几何,可以作为 AND 逻辑门.
结论:
- 开发的自抑制设计范式为创建高度敏感,遗传编码的开启蛋白酶生物传感器提供了强大的方法.
- 这一策略为控制分裂蛋白的活性提供了一个可通用的方法,在诊断和药物发现方面有潜在的应用.
更多相关视频
10:33Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors
Published on: October 26, 2015
19:16The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis
Published on: March 17, 2010
相关概念视频
Single-Strand DNA Binding Proteins
For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
Protein Folding
Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
Protein Folding
Overview
Molecular Chaperones and Protein Folding
The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
The...
The...
