Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Russia.
The Branch of the Institute of Bioorganic Chemistry, Russian Academy of Sciences, 142290 Pushchino, Russia.
Int J Mol Sci. 2021 Jul 7;22(14):7291. doi: 10.3390/ijms22147291.
Bacterial S1 protein is a functionally important ribosomal protein. It is a part of the 30S ribosomal subunit and is also able to interact with mRNA and tmRNA. An important feature of the S1 protein family is a strong tendency towards aggregation. To study the amyloidogenic properties of S1, we isolated and purified the recombinant ribosomal S1 protein of . Using the FoldAmyloid, Waltz, Pasta 2.0, and AGGRESCAN programs, amyloidogenic regions of the protein were predicted, which play a key role in its aggregation. The method of limited proteolysis in combination with high performance liquid chromatography and mass spectrometric analysis of the products, made it possible to identify regions of the S1 protein from that are protected from the action of proteinase K, trypsin, and chymotrypsin. Sequences of theoretically predicted and experimentally identified amyloidogenic regions were used to synthesize four peptides, three of which demonstrated the ability to form amyloid-like fibrils, as shown by electron microscopy and fluorescence spectroscopy. The identified amyloidogenic sites can further serve as a basis for the development of new antibacterial peptides against the pathogenic microorganism .
细菌 S1 蛋白是一种具有重要功能的核糖体蛋白。它是 30S 核糖体亚基的一部分，也能够与 mRNA 和 tmRNA 相互作用。S1 蛋白家族的一个重要特征是强烈的聚集倾向。为了研究 S1 的淀粉样特性，我们分离并纯化了 的重组核糖体 S1 蛋白。使用 FoldAmyloid、Waltz、Pasta 2.0 和 AGGRESCAN 程序，预测了该蛋白的淀粉样区域，这些区域在其聚集过程中起着关键作用。采用有限蛋白酶解方法结合高效液相色谱和产物的质谱分析，确定了 S1 蛋白的几个区域对蛋白酶 K、胰蛋白酶和糜蛋白酶的作用具有保护作用。理论预测和实验鉴定的淀粉样区域序列被用来合成四个肽，其中三个肽表现出形成类似淀粉样纤维的能力，这可以通过电子显微镜和荧光光谱来证明。鉴定出的淀粉样形成部位可以进一步作为开发针对致病微生物 的新型抗菌肽的基础。
