Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, 160 00 Prague, Czech Republic; Second Faculty of Medicine, Charles University, 150 06 Prague, Czech Republic.
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, 160 00 Prague, Czech Republic.
Int J Biol Macromol. 2021 Jan 31;168:1-12. doi: 10.1016/j.ijbiomac.2020.11.216. Epub 2020 Dec 5.
Constantly increasing attention to bioengineered proteins has led to the rapid development of new functional targets. Here we present the biophysical and functional characteristics of the newly designed CaM/AMBN-Ct fusion protein. The two-domain artificial target consists of calmodulin (CaM) and ameloblastin C-terminus (AMBN-Ct). CaM as a well-characterized calcium ions (Ca) binding protein offers plenty of options in terms of Ca detection in biomedicine and biotechnologies. Highly negatively charged AMBN-Ct belongs to intrinsically disordered proteins (IDPs). CaM/AMBN-Ct was designed to open new ways of communication synergies between the domains with potential functional improvement. The character and function of CaM/AMBN-Ct were explored by biophysical and molecular modelling methods. Experimental studies have revealed increased stability and preserved CaM/AMBN-Ct function. The results of molecular dynamic simulations (MDs) outlined different interface patterns between the domains with potential allosteric communication within the fusion.
人们对生物工程蛋白的关注度不断提高，这促使新的功能靶点迅速发展。在这里，我们介绍了新设计的 CaM/AMBN-Ct 融合蛋白的生物物理和功能特性。该双域人工靶标由钙调蛋白 (CaM) 和釉原蛋白 C 端 (AMBN-Ct) 组成。CaM 作为一种具有良好特性的钙离子 (Ca) 结合蛋白，在生物医学和生物技术中的 Ca 检测方面提供了多种选择。带负电荷的 AMBN-Ct 高度属于无规卷曲蛋白 (IDPs)。CaM/AMBN-Ct 的设计旨在为域间协同作用开辟新的沟通途径，具有潜在的功能改善。通过生物物理和分子建模方法研究了 CaM/AMBN-Ct 的特性和功能。实验研究表明，融合蛋白的稳定性增加，且保留了 CaM/AMBN-Ct 的功能。分子动力学模拟 (MD) 的结果描绘了融合蛋白中域间不同的界面模式，存在潜在的变构通讯。
