9.4 Tesla Fourier Transform Mass Spectrometer

描述（由申请人提供）：该资助的目的是购买FT-MS仪器，以扩大UAB在三个特定领域的质谱分析能力：（1）使用H-D交换来探测蛋白质结构和动力学，（2）识别蛋白质修饰（永久和瞬时），以及（3）分析组织间质液的蛋白质和肽含量。FT-MS提供的高质量分辨率，即使是大的蛋白质分子，将提供重要的新功能，蛋白质折叠的研究使用H-D交换。UAB的研究重点是病毒和噬菌体蛋白的组装。虽然H-D交换已经被用于使用MALDI-TOF质谱法研究HLV病毒的衣壳蛋白，但是使用该技术的蛋白质的覆盖是有限的。在佛罗里达州立大学的国家高场磁体设施中使用Fr-MS获得的初步数据已经证明了FT-MS在鉴定通过胃蛋白酶消化产生的肽和所获得的数据的高再现性方面的能力。FT-MS的第二个重要用途是对经过生化或化学修饰的肽进行测序。虽然我们有一个Q-tof 2仪器，提供高分辨率的MS-MS数据，准确的母离子的选择是不可能的四极过滤器。Fr-MS仪器还将提供在完整蛋白质水平上检查蛋白质修饰的能力。这对酶-底物-产物复合物的研究具有重要意义。这种方法对于经历乒乓反应机制的酶特别有用，其中底物形成共价酶中间体。再次，Fr-MS获得高分辨率、高灵敏度的质谱数据的能力对于组织间液成分的研究也非常有价值。这种物质是细胞所沐浴的物质，含有许多信号分子（细胞因子，旁分泌生长因子等）。由于研究者可获得的量很小，除了大脑，对脑脊液的研究在很大程度上被忽视了。描述单个细胞的局部环境对于理解疾病和设计对抗疾病的药物至关重要。总的来说，FT-MS仪器将维持高质量的生物医学研究，这是UAB的标志。

DESCRIPTION (provided by applicant): The objective of this grant is to purchase a FT-MS instrument to expand the mass spectrometry capabilities at UAB in three specific areas: (1) use of H-D exchange to probe protein structure and dynamics, (2) to identify protein modifications (permanent and transient), and (3) to analyze the protein and peptide content of tissue interstitial fluids. The high mass resolution offered by FT-MS. even for large protein molecules, will provide important new capabilities for the study of protein folding using H-D exchange. A focus of research at UAB is in the assembly of virus and phage proteins. Although H-D exchange has been used to study the capsid protein of the HLV virus using MALDI-TOF mass spectrometry, the coverage of the protein with this technique is limited. Preliminary data obtained using Fr-MS at the National High Field Magnet Facility at Florida State University have demonstrated the power of the FT-MS in both the identification of the peptides produced by digestion with pepsin and the high reproducibility of the data obtained. A second important use of FT-MS is in sequencing of peptides that have undergone biochemical or chemical modification. Although we have a Q-tof2 instrument that gives high resolution MS-MS data, accurate selection of the parent ion is not possible with a quadrupole filter. The Fr-MS instrument will also provide the ability to examine protein modification at the intact protein level. This will be valuable for the study of enzyme-substrate-product complexes. This approach is particularly useful for enzymes undergoing a Ping-Pong reaction mechanism where the substrate forms a covalent enzyme intermediate. Thirdly, the ability of Fr-MS to obtain high resolution mass spectral data and at high sensitivity is also very valuable for the study of the components of interstitial fluids. This material is what cells are bathed in and contains many signaling molecules (cytokines, paracrine growth factors etc). Because of the tiny amounts available to the investigator, except for the brain, study of interstital fluids has been largely ignored. Describing the local environment of individual cells is essential for understanding diseases and for the design of drugs to combat them. Overall, the FT-MS instrument will sustain the high quality of biomedical research that is a hallmark of UAB.