Fast and Angström-resolution AFM to visualise conformational change in biomolecules

快速且埃级分辨率的 AFM 可可视化生物分子的构象变化

• 批准号: BB/G011729/1
• 负责人: Bart Hoogenboom
• 金额: $ 42.1万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2009
• 资助国家: 英国
• 起止时间: 2009 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG011729%2F1
• 关键词: Fast; Angstr; resolution; AFM; visualise

Atomic Force Microscopy (AFM) basically acts as a miniature blind man's stick ('cantilever') following the contours of a sample surface, and line by line reconstructing a three-dimensional representation of the surface topography. This line-by-line scanning is a fundamental difference from other, more common microscopy techniques and a main reason why it generally takes minutes to complete a single image. AFM is unique in combining sub-molecular resolution imaging with the ability to operate in liquids. For high-resolution imaging of biological samples, molecules are generally adsorbed on a hard surface, which is the only compromise compared to physiological conditions. Membrane proteins are samples of particular interest, since they represent more than 50% of modern drug targets and therefore are of major pharmaceutical importance. Their function as molecular nanomachines is determined by Angstrom-sized structural ('conformational') changes occurring at millisecond time scales. For applications in future healthcare and for basic scientific understanding, the crucial question is how molecular structure and changes in this structure relate to the biological function of membrane proteins. This project combines high-resolution AFM techniques (that have yielded atomic resolution!) with fast scanning, to obtain images of membrane proteins with Angstrom spatial and millisecond temporal resolution. This will enable us to visualise conformational changes in real time and observe biomolecules at work. This will be demonstrated on bacteriorhodopsin, a light-driven molecular machine that pumps protons through the cell membrane.

原子力显微镜(AFM)基本上就是一个跟随样品表面轮廓的微型盲人手杖(悬臂)，逐线重建表面形貌的三维表示。这种逐行扫描是与其他更常见的显微镜技术的根本区别，也是通常需要几分钟才能完成一张图像的主要原因。原子力显微镜在结合亚分子分辨率成像和在液体中操作的能力方面是独一无二的。对于生物样品的高分辨率成像，分子通常吸附在坚硬的表面上，这是与生理条件相比唯一的折衷。膜蛋白是特别令人感兴趣的样品，因为它们代表了现代药物靶标的50%以上，因此具有重大的药学意义。它们作为分子纳米机器的功能是由毫秒时间尺度上发生的埃大小的结构(构象)变化决定的。对于在未来的医疗保健中的应用和对基础科学的理解，关键的问题是分子结构和这种结构的变化如何与膜蛋白的生物学功能相关。这个项目结合了高分辨率的AFM技术(已经产生了原子分辨率！)通过快速扫描，获得Angstrom空间分辨率和毫秒时间分辨率的膜蛋白图像。这将使我们能够实时可视化构象变化，并观察工作中的生物分子。这将在细菌视紫红质上得到证明，这是一种光驱动的分子机器，可以将质子泵过细胞膜。

项目成果

Nanoscale stiffness topography reveals structure and mechanics of the transport barrier in intact nuclear pore complexes.

• DOI: 10.1038/nnano.2014.262
• 发表时间: 2015-01
• 期刊: Nature nanotechnology
• 影响因子: 38.3

Resolving the structure of a model hydrophobic surface: DODAB monolayers on mica

• DOI: 10.1039/c2ra20108a
• 发表时间: 2012-01-01
• 期刊: RSC ADVANCES
• 影响因子: 3.9
• 作者: Gosvami, Nitya Nand;Parsons, Edward;Perkin, Susan
• 通讯作者: Perkin, Susan

Improved Kelvin probe force microscopy for imaging individual DNA molecules on insulating surfaces

• DOI: 10.1063/1.3512867
• 发表时间: 2010-11-15
• 期刊: APPLIED PHYSICS LETTERS
• 影响因子: 4
• 作者: Leung, Carl;Maradan, Dario;Hoogenboom, Bart W.
• 通讯作者: Hoogenboom, Bart W.

Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin.

• DOI: 10.7554/elife.04247
• 发表时间: 2014-12-02
• 期刊: eLife
• 影响因子: 7.7
• 作者: Leung C;Dudkina NV;Lukoyanova N;Hodel AW;Farabella I;Pandurangan AP;Jahan N;Pires Damaso M;Osmanović D;Reboul CF;Dunstone MA;Andrew PW;Lonnen R;Topf M;Saibil HR;Hoogenboom BW
• 通讯作者: Hoogenboom BW

Enhanced quality factors and force sensitivity by attaching magnetic beads to cantilevers for atomic force microscopy in liquid

• DOI: 10.1063/1.4768713
• 发表时间: 2012-11
• 期刊: Journal of Applied Physics
• 影响因子: 3.2
• 作者: S. Hoof;N. Gosvami;B. Hoogenboom