病毒的原子力显微镜:稳定性,分解和基因组释放
Miguel Cantero1, María Jesús Rodríguez-Espinosa1,2, Klara Strobl1
1Departamento de Física de la Materia Condensada, Universidad Autónoma de Madrid, Madrid, Spain.
Methods in molecular biology (Clifton, N.J.)
|October 12, 2023
概括
原子力显微镜 (AFM) 图像和操纵单个蛋白质外,如病毒,在液体中. 该技术评估病毒特性,货物释放,衰老和对内部蛋白质的拥挤影响.
科学领域:
- 生物物理学的生物物理.
- 纳米技术纳米技术
- 结构生物学 结构生物学
背景情况:
- 原子力显微镜 (AFM) 提供液体中的生物标本的纳米分辨率成像.
- AFM探测器,包括一个微型杆上的纳米尖端,与样品机械相互作用.
- 蛋白质外,包括病毒,容易接受基于AFM的成像和操纵.
研究的目的:
- 对吸附蛋白质外到表面进行AFM分析的方法进行审查.
- 为了研究尖端几何学对AFM地形学的影响.
- 探索AFM在不同的液体条件下监测传染性胃肠炎病毒 (TGEV) 的能力.
- 详细介绍AFM研究病毒载荷释放,老化和多层结构的方法,使用缩和疲劳分析.
- 提出一项联合的AFM和光学研究,研究P22菌体囊中的拥挤效应.
主要方法:
- 蛋白质外的表面吸附协议.
- 使用不同的尖端几何形状的AFM成像.
- 对于TGEV监控的环境控制.
- 单和疲劳测定用于病毒分析.
- 结合了AFM和光显微镜.
主要成果:
- 建立了蛋白质外吸附的协议,并讨论了AFM地形学上的尖端几何效应.
- 证明了AFM在不同液态环境下监测TGEV的实用性.
- 描述病毒载荷释放,衰老动态和多层病毒的机械性质.
- 量化了宏分子拥挤对P22细菌囊中的封装蛋白质 (GFP) 的影响.
结论:
- AFM是一种多功能工具,用于成像,操纵和表征液体中的单个蛋白质.
- AFM能够对病毒行为的详细研究,包括环境反应,衰老和内部动态.
- 结合AFM和光显微镜,可以对像拥挤的病毒囊体这样的复杂生物系统提供强大的洞察力.
相关概念视频
Viral Structure
62.4K
Viruses are extraordinarily diverse in shape and size, but they all have several structural features in common. All viruses have a core that contains a DNA- or RNA-based genome. The core is surrounded by a protective coat of proteins called the capsid. The capsid is composed of subunits called capsomeres. The capsid and genome-containing core are together known as the nucleocapsid.
62.4K
Studying the Cytoskeleton
6.3K
The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
6.3K
Cryo-electron Microscopy
3.4K
Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
3.4K
Atomic Force Microscopy
3.4K
Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...
3.4K


