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

基本信息

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

项目摘要

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) 基本上充当微型盲人棍（“悬臂”），跟随样品表面的轮廓，并逐行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单个图像的主要原因。 AFM 的独特之处在于将亚分子分辨率成像与在液体中操作的能力相结合。对于生物样品的高分辨率成像，分子通常吸附在硬表面上，这是与生理条件相比的唯一妥协。膜蛋白是人们特别感兴趣的样品，因为它们代表了超过 50% 的现代药物靶标，因此具有重要的制药重要性。它们作为分子纳米机器的功能是由毫秒级发生的埃大小的结构（“构象”）变化决定的。对于未来医疗保健的应用和基础科学理解，关键问题是分子结构和该结构的变化如何与膜蛋白的生物功能相关。该项目将高分辨率 AFM 技术（已产生原子分辨率！）与快速扫描相结合，以获得具有埃空间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，细菌视紫红质是一种光驱动的分子机器，可以通过细胞膜泵送质子。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
作者：
通讯作者：

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