A mass spectrometer for protein hydrogen exchange studies

描述(由申请者提供)：申请的资金旨在建造一个特殊用途的设施，以支持利用氢交换-质谱学(HX/MS)技术研究高级蛋白质结构-功能问题。在过去的十年里，HX方法和多维核磁共振分析一起被越来越多地用于提供关于相对较小的蛋白质的结构、结构变化、能量学、动力学和折叠的详细的氨基酸分解信息。现在很清楚，这些重要的能力可以通过结合MS分析的蛋白质分解方法扩展到更大、更有趣的蛋白质。八个主要用户希望将这些能力应用于他们正在进行的以下研究：淀粉样蛋白和前淀粉样蛋白的结构和性质；着丝粒的组蛋白结构及其在染色体分离中的作用；尿激酶及其受体的相互作用；甲型流感病毒M2膜通道蛋白的结构和药物相互作用；载脂蛋白及其脂运输复合体的结构和功能；免疫球蛋白轻链淀粉样蛋白的形成机制；钙调蛋白的动力学、熵和功能。通过氢交换方法，这种关于这种困难的生物分子系统的基本生物物理信息是唯一可以获得的。在最近的工作中，已经证明了一种系统配置，它将使多个组能够轻松地共享MS设施。该设施将支持主要用户的工作，并将有助于在这个园区和更广泛的范围内形成新兴和强大的HX/MS技术的核心。宾夕法尼亚大学承诺为该设施的建设提供大量帮助，并继续提供资金支持。 所要求的设施将组装一项基于蛋白质氢交换和质谱分析的新兴技术的强大能力。这一能力将使一系列旨在了解和控制蛋白质分子在健康和疾病中的性质和相互作用的研究工作成为可能。

DESCRIPTION (provided by applicant): The funding requested here is intended to construct a special purpose facility to support the investigation of advanced protein structure-function problems by the use of hydrogen exchange - mass spectrometry (HX/MS) technology. Over the last decade HX methods together with multi- dimensional NMR analysis have been increasingly used to provide detailed amino acid-resolved information on structure, structure change, energetics, dynamics, and folding for relatively small proteins. It is now clear that these important capabilities can be extended to larger more interesting proteins by use of a proteolytic fragmentation method together with MS analysis. The eight major users want to apply these capabilities to their ongoing studies on: amyloid and pre-amyloid structure and properties; centromeric histone structure and its role in chromosome segregation; urokinase and its receptor interactions; structure and drug interactions in the M2 membrane channel protein of influenza A virus; the structure and function of apolipoproteins and their lipid transport complexes; the mechanism of immunoglobin light chain amyloid formation; calmodulin dynamics, entropy and function. This kind of fundamental biophysical information on such difficult biomolecular systems is uniquely accessible to hydrogen exchange approaches. A system configuration that will make it easily possible for multiple groups to share the MS facility has been demonstrated in recent work. The facility will support the work of the major users and will help to nucleate the emerging and powerful HX/MS technology on this campus and more broadly. The University of Pennsylvania has committed substantial help to construct the facility and support it with continuing funding. The facility requested will assemble the powerful capabilities of a newly emerging technology based on protein hydrogen exchange and mass spectrometric analysis. This capability will make possible a range of research efforts directed at understanding and controlling the properties and interactions of protein molecules in health and disease.