相关实验视频
Updated: Jun 4, 2025

05:49
Protein Engineering by Yeast Surface Display
Published on: November 29, 2024
1.0K
以进化为基础的蛋白质工程:在transthyretins和5-hydroxyisourate hydrolases之间进行功能切换
Rafael Pereira Lemos1,2, Julia T Rodrigues1, Gabriel Portwood1
1Laboratory for Macromolecular Biophysics - LBM, Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, Brazil.
Journal of biomolecular structure & dynamics
|December 20, 2024
概括
研究人员通过改变跨甲基素 (TTR) 和5-基苏酸酶 (HIUase) 序列来设计新的酶. 这种以进化为基础的方法成功地创造了具有改变功能的活性酶,证明了残留物特定的工程能力.
科学领域:
- 生物化学 生物化学
- 进化生物学 进化生物学
- 蛋白质工程是指蛋白质的工程.
背景情况:
- 甲状腺激素 (TTR) 是一种脊椎动物的运输蛋白质,对甲状腺激素的分发至关重要.
- TTR是从编码5-氧苏酸酸酶 (HIUase) 的基因进化而来的,这是一种参与尿酸代谢的酶.
- HIUase是普遍存在的,而TTR是脊椎动物特异的;两者都形成具有相似结构的同位基.
研究的目的:
- 通过计算蛋白质工程来研究HIUase和TTR函数相互转换的潜力.
- 探索一种以进化为基础的方法,通过利用保存的残留物来修改蛋白质功能.
主要方法:
- 通过在HIUase和TTR代表之间替换相关的局部保存位置来设计新的蛋白质序列.
- 利用计算建模来改进,验证和分析结构性质 (腔体积,几何,聚合倾向,静电学).
- 进行了分子动力学模拟,以评估工程突变物在绑定和不绑定状态中的稳定性.
主要成果:
- 计算分析揭示了工程蛋白质及其连接体之间的腔体和几何体积的差异.
- 分子动力学模拟证实了束状态突变复合物的稳定性.
- 酶分析表明,这些工程突变物具有活跃的新型酶功能.
结论:
- 基于进化的蛋白质工程策略有效地创造了具有改变性质的功能性酶.
- 这种方法允许对残留物进行特定修改,突出显示HIUase和TTR等进化相关蛋白质中的保存残留物.
- 这项研究为设计基于进化关系的定制功能蛋白质提供了一个框架.
相关概念视频
Allosteric Proteins-ATCase
5.7K
Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
5.7K
Protein Complexes with Interchangeable Parts
2.5K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.5K
Ligand Binding and Linkage
4.8K
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
4.8K
Covalently Linked Protein Regulators
6.8K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
These groups modify specific amino acids in a protein....
6.8K
The Unfolded Protein Response
4.4K
The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...
4.4K
Protein Modifications in the RER
5.0K
Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal...
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal...
5.0K

