{"title":"Structural Investigations of Cargo Molecules Inside Icosahedrally Symmetric Encapsulin by VUVCD Spectroscopic Measurements","authors":"Shiori Kumamoto, Akima Yamamoto, Yu Shiratsuchi, Koichi Matsuo, Akifumi Higashiura, Daisuke Hira","doi":"10.1002/chir.23700","DOIUrl":null,"url":null,"abstract":"<p>Prokaryotes organize intracellular compartments with protein-based organelles called encapsulins. Encapsulins with icosahedral symmetry can encapsulate specific cargo proteins mediated by targeting peptides or encapsulation-mediating domains. Encapsulins have been used in eukaryotic cells for bioengineering, vaccine development, and nanoparticle alignment. Their versatility makes them attractive for research; however, detailed structural information on encapsulins is crucial for further applied research. However, cargo proteins are randomly oriented inside the icosahedral encapsulins. The random orientation of cargo proteins presents a challenge for structural analysis that relies on averaging processes such as x-ray crystallography and cryo-electron microscopy (cryo-EM) single-particle imaging. Therefore, we aimed to accurately estimate the secondary structure content and elucidate the structure of cargo proteins inside the particle by measuring the circular dichroism (CD) spectra using vacuum ultraviolet circular dichroism (VUVCD) spectroscopy. Thus, the structure of the cargo protein inside encapsulin was evaluated. This approach could potentially set a standard for evaluating cargo proteins inside particles in future applied research on encapsulins.</p>","PeriodicalId":10170,"journal":{"name":"Chirality","volume":"36 8","pages":""},"PeriodicalIF":2.8000,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/chir.23700","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chirality","FirstCategoryId":"92","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/chir.23700","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
Abstract
Prokaryotes organize intracellular compartments with protein-based organelles called encapsulins. Encapsulins with icosahedral symmetry can encapsulate specific cargo proteins mediated by targeting peptides or encapsulation-mediating domains. Encapsulins have been used in eukaryotic cells for bioengineering, vaccine development, and nanoparticle alignment. Their versatility makes them attractive for research; however, detailed structural information on encapsulins is crucial for further applied research. However, cargo proteins are randomly oriented inside the icosahedral encapsulins. The random orientation of cargo proteins presents a challenge for structural analysis that relies on averaging processes such as x-ray crystallography and cryo-electron microscopy (cryo-EM) single-particle imaging. Therefore, we aimed to accurately estimate the secondary structure content and elucidate the structure of cargo proteins inside the particle by measuring the circular dichroism (CD) spectra using vacuum ultraviolet circular dichroism (VUVCD) spectroscopy. Thus, the structure of the cargo protein inside encapsulin was evaluated. This approach could potentially set a standard for evaluating cargo proteins inside particles in future applied research on encapsulins.
原核生物利用基于蛋白质的细胞器--封装体--来组织细胞内隔室。具有二十面体对称性的封装蛋白可通过靶向肽或封装介导结构域封装特定的货物蛋白。封装蛋白已在真核细胞中用于生物工程、疫苗开发和纳米粒子排列。封装蛋白的多功能性使其对研究具有吸引力;然而,封装蛋白的详细结构信息对进一步的应用研究至关重要。然而,货物蛋白在二十面体封装体内是随机定向的。货物蛋白的随机取向给依赖于 X 射线晶体学和低温电子显微镜(cryo-EM)单颗粒成像等平均过程的结构分析带来了挑战。因此,我们利用真空紫外圆二色性光谱(VUVCD)测量圆二色性光谱,旨在准确估算二级结构含量,并阐明颗粒内货物蛋白的结构。因此,包囊蛋白内部的货物蛋白结构得到了评估。这种方法有可能在未来的封装蛋白应用研究中为评估颗粒内的货物蛋白设定一个标准。
期刊介绍:
The main aim of the journal is to publish original contributions of scientific work on the role of chirality in chemistry and biochemistry in respect to biological, chemical, materials, pharmacological, spectroscopic and physical properties.
Papers on the chemistry (physiochemical, preparative synthetic, and analytical), physics, pharmacology, clinical pharmacology, toxicology, and other biological aspects of chiral molecules will be published.