评估His标签对稳基酶变异的活性和稳定性的影响
Monika Štulajterová1, Ľuboš Ambro2, Dagmar Sedláková3
1Department of Biophysics, Faculty of Science, P. J. Šafárik University in Košice, Jesenná 5, 04154, Košice, Slovakia.
International journal of biological macromolecules
|September 14, 2025
概括
在酸性条件下,N终端多胺 (His-tag) 显著破坏稳定性,影响其治疗潜力. 萨克星展出了最强大的性能,使其成为进一步开发的有希望的候选人.
科学领域:
- 生物化学 生物化学
- 蛋白质工程是指蛋白质工程.
- 生物物理学的生物物理.
背景情况:
- 稳氨基酶 (SAK) 是一种具有血栓解压潜力的强效等离子体激活剂.
- 由于免疫性和稳定性问题,SAK的临床使用受到阻碍.
- 诸如多胺 (His-tag) 这样的亲和标签对SAK特性的影响尚不清楚.
研究的目的:
- 调查His标记对SAK变种稳定性和活性的影响.
- 为了比较不同SAK类型 (STAR和42D) 的生物物理和功能性质,有或没有His标签.
- 为了确定最适合治疗开发的SAK变体.
主要方法:
- 对四种SAK变体进行系统的调查:SAK STAR和SAK 42D的野生型和三重氨酸 (3A) 形式.
- 使用循环二重化和托芬光谱学评估热和pH稳定性.
- 通过染色体测试评估等离子体激活效率.
主要成果:
- 他的标记对热稳定性的影响很小,对功能活动的影响也很小.
- 在酸性条件下,他的标签显著破坏了SAK的稳定性,改变了展开的过渡.
- 与SAK 42D 3A相比,SAK STAR表现出优越的结构和功能强度,SAK 42D 3A容易聚合.
结论:
- 亲和标签可以显著改变稳基酶的生物物理特性,特别是在酸性条件下.
- SAK STAR被确定为最稳定和功能最强大的变种.
- 萨克斯塔是未来血栓溶解治疗开发中最有前途的候选药物.
相关概念视频
01:24Tagging and Fusion Proteins
8.3K
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...
8.3K
00:49Ligand Binding and Linkage
5.5K
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...
5.5K
01:19Allosteric Proteins-ATCase
6.5K
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...
6.5K