对旧蛋白质的新技巧:非酶蛋白中的单个突变导致各种酶活动
Yurii S Moroz1, Tiffany T Dunston1, Olga V Makhlynets1
1Department of Chemistry, Syracuse University , 111 College Place, Syracuse, New York 13244, United States.
Journal of the American Chemical Society
|November 12, 2015
概括
研究人员通过将单个突变引入一个小蛋白质, 制造出一种高效的雌激酶AlleyCatE. 这一突破表明, 微小的变化可以释放重要的催化功能,
科学领域:
- 生物化学
- 蛋白质工程
- 酶催化
背景情况:
- 了解酶活性对于蛋白质设计至关重要.
- 目前的蛋白质设计方法可能复杂且计算密集.
研究的目的:
- 在计算上开发一种廉价的,极简的方法来引入蛋白质中的催化功能.
- 从非酶蛋白域中设计一个高效的酶.
主要方法:
- 使用简单的设计策略,专注于单一的反应残留物.
- 将卡尔莫杜林的C终端域转化为雌激酶 (AlleyCatE).
- 评估了催化效率,立体选择性和性调节.
主要成果:
- 成功将calmodulin转化为一种高效的立体选择性雌激酶 (AlleyCatE).
- 与之前设计的 de novo esterases 相比,AlleyCatE 的催化效率更高.
- 设计的蛋白质在水解,Kemp消除和逆醇反应中表现出催化活性.
结论:
- 一个单一的突变可以为小的非酶蛋白提供有效的催化活性.
- 最简单的设计协议扩展了新型蛋白质设计的能力.
- 蛋白质可以通过向突变作为后续进化途径的入口点.
更多相关视频
11:36A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
Published on: July 3, 2016
11.4K
11:49A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
Published on: August 21, 2018
7.0K
相关概念视频
Mutations
96.4K
Overview
96.4K
Mutations
45.5K
Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations can be characterized in a number of different ways: whether and how they alter the amino acid sequence of the protein, whether they occur over a small or large area of DNA, and whether they occur in somatic cells or germline cells.
Chromosomal Alterations Are Large-Scale Mutations
While point mutations are changes in a single nucleotide in...
Chromosomal Alterations Are Large-Scale Mutations
While point mutations are changes in a single nucleotide in...
45.5K
Mutations
13.7K
13.7K
Mutations in Microorganisms
1.1K
Mutations are heritable changes in an organism’s genome involving alterations in the base sequence of DNA or RNA. These changes can influence cellular processes and phenotypic traits, potentially transforming the unaltered wild type into a mutant form. Such changes, termed forward mutations, are pivotal in shaping the genetic diversity of organisms.RNA viruses exhibit the highest mutation rates due to the absence of robust proofreading mechanisms during genome replication. In contrast,...
1.1K
Point and Frameshift Mutations
1.6K
Point mutations are genetic alterations involving the change of a single nucleotide base pair in DNA. Depending on how the alteration affects protein synthesis, they can lead to various consequences.Point mutations fall into the following types:Silent mutations occur when a nucleotide change does not alter the amino acid sequence due to the redundancy of the genetic code. For instance, changing ACC to ACA still encodes threonine, leaving the protein function unaffected. This occurs because...
1.6K
Spontaneous and Induced Mutations
3.0K
Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
3.0K
