Enhanced Mass Spectrometry Facilities for the Astbury Centre for Structural Molecular Biology

阿斯特伯里结构分子生物学中心的增强型质谱设施

• 批准号: BB/E012558/1
• 负责人: Alison Ashcroft
• 金额: $ 30.04万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE012558%2F1
• 关键词: Enhanced; Mass; Spectrometry; Facilities; Astbury

Mass spectrometry is an analytical technique whereby biological samples such as peptides, proteins and nucleotides can be mass measured. Even fragile macromolecular complexes consisting of multiple protein molecules and other species such as RNA or DNA molecules which are held together by weak, non-covalent interactions, can be preserved and mass measured, if carefully controlled conditions are chosen. Some mass spectrometers also allow fragmentation of the sample to take place during the analysis. This generates a number of fragments each of which is mass measured. The results from these fragments can then be stitched together, like pieces of a jigsaw puzzle, to give a picture of the structural composition of the sample. This is particularly useful for peptides, whose amino acid sequence can be determined in this way. We propose to replace two of our old mass spectrometers with a state-of-the-art instrument that will enable us to both measure masses and carry out structural determinations. This equipment is vital for our research, which covers many aspects of structural molecular biology. One project involves an investigation into the RNA-triggered assembly of virus capsids from protein monomers. By studying this reaction and characterising key intermediates on the way to the final virus capsid, we can seek to understand, and subsequently devise methods to prevent, virus assembly. Another project looks into the way certain proteins fold into their unique 3D shape. If a protein unfolds, there is an opportunity for it to fold incorrectly. This phenomenon is thought to occur in vivo, where some ~20 proteins misfold and aggregate into large, insoluble, polymeric complexes which are associated with many incurable diseases such as Alzheimer's, CJD, and BSE. Other projects we are carrying out include studying the structure and behaviour of proteins and peptides from different organisms such as the mosquito and fruit fly, as well as plants, in order to determine how these species function. Peptide sequencing will be vital for these studies. The new mass spectrometer will be used by many people in the Astbury Centre for Structural Molecular Biology and the Faculty of Biological Sciences, as well as our collaborators in the Schools of Chemistry, Physics, Medicine and Dentistry: thus it will bring together many scientific disciplines. Not only will it enhance our research, which is aimed directly at understanding how biological systems work in order to improve the quality of life, it will also be used to train post-graduate and post-doctoral research workers from many groups, and will provide us with an opportunity to develop new mass spectrometric techniques that we will apply to novel bioanalyses and these will be beneficial to many other people in academia and in industry.

质谱法是一种分析技术，通过它可以对生物样品如肽、蛋白质和核苷酸进行质量测量。即使是由多个蛋白质分子和其他物质（如RNA或DNA分子）组成的脆弱的大分子复合物，如果选择了精心控制的条件，也可以通过弱的非共价相互作用保持在一起并进行质量测量。一些质谱仪还允许在分析期间发生样品的碎片化。这产生了许多碎片，每个碎片都被质量测量。这些碎片的结果可以像拼图一样拼接在一起，以给出样品结构组成的图片。这对于肽特别有用，其氨基酸序列可以以这种方式确定。我们建议用最先进的仪器取代两台旧的质谱仪，使我们能够测量质量并进行结构测定。这些设备对我们的研究至关重要，因为我们的研究涵盖了结构分子生物学的许多方面。其中一个项目涉及对RNA触发的蛋白单体组装病毒衣壳的研究。通过研究这个反应，并对最终病毒衣壳形成过程中的关键中间体进行表征，我们可以寻求理解，并随后设计出防止病毒组装的方法。另一个项目研究了某些蛋白质折叠成其独特的3D形状的方式。如果蛋白质展开，它就有可能错误地折叠。这种现象被认为发生在体内，其中约20种蛋白质错误折叠并聚集成大的不溶性聚合物复合物，其与许多不治之症如阿尔茨海默氏症、CJD和BSE相关。我们正在开展的其他项目包括研究来自不同生物体（如蚊子和果蝇）以及植物的蛋白质和肽的结构和行为，以确定这些物种的功能。肽序列测定对这些研究至关重要。新的质谱仪将被许多人在阿斯特伯里中心的结构分子生物学和生物科学学院，以及我们的合作者在化学，物理，医学和牙科学院使用：因此，它将汇集许多科学学科。它不仅将加强我们的研究，这是直接旨在了解生物系统如何工作，以提高生活质量，它也将用于培训研究生和博士后研究工作者从许多群体，并将为我们提供一个机会，开发新的质谱技术，我们将应用于新的生物分析，这将有利于许多其他人在学术界和行业

项目成果

Investigating the Structural Compaction of Biomolecules Upon Transition to the Gas-Phase Using ESI-TWIMS-MS.

• DOI: 10.1007/s13361-017-1689-9
• 发表时间: 2017-09
• 期刊: Journal of the American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: Devine PWA;Fisher HC;Calabrese AN;Whelan F;Higazi DR;Potts JR;Lowe DC;Radford SE;Ashcroft AE
• 通讯作者: Ashcroft AE

Using hydroxyl radical footprinting to explore the free energy landscape of protein folding.

• DOI: 10.1016/j.ymeth.2015.02.018
• 发表时间: 2015-11-01
• 期刊: Methods (San Diego, Calif.)
• 作者: Calabrese AN;Ault JR;Radford SE;Ashcroft AE
• 通讯作者: Ashcroft AE

Nucleocapsid protein structures from orthobunyaviruses reveal insight into ribonucleoprotein architecture and RNA polymerization.

• DOI: 10.1093/nar/gkt268
• 发表时间: 2013-06
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Ariza A;Tanner SJ;Walter CT;Dent KC;Shepherd DA;Wu W;Matthews SV;Hiscox JA;Green TJ;Luo M;Elliott RM;Fooks AR;Ashcroft AE;Stonehouse NJ;Ranson NA;Barr JN;Edwards TA
• 通讯作者: Edwards TA