A mass spectrometer for protein hydrogen exchange studies

描述（由申请人提供）：申请的资金用于建设一个特殊用途的设施，以支持使用氢交换-质谱（HX/MS）技术研究高级蛋白质结构-功能问题。在过去的十年中，HX方法与多维核磁共振分析一起越来越多地用于提供有关相对较小蛋白质的结构，结构变化，能量学，动力学和折叠的详细氨基酸解析信息。现在很清楚，这些重要的能力可以扩展到更大的更有趣的蛋白质，通过使用蛋白质裂解裂解方法和质谱分析。八个主要用户希望将这些功能应用于他们正在进行的研究：淀粉样蛋白和前淀粉样蛋白结构和性质；着丝粒组蛋白结构及其在染色体分离中的作用尿激酶及其受体相互作用；甲型流感病毒M2膜通道蛋白结构及药物相互作用载脂蛋白及其脂转运复合物的结构和功能；免疫球蛋白轻链淀粉样蛋白的形成机制钙调素动力学、熵和功能。这种复杂的生物分子系统的基本生物物理信息只有通过氢交换方法才能获得。在最近的工作中已经演示了一种系统配置，它可以使多个组轻松地共享MS设施。该设施将支持主要用户的工作，并将有助于在这个校园和更广泛的范围内形成新兴和强大的HX/MS技术。宾夕法尼亚大学承诺为建设该设施提供大量帮助，并提供持续的资金支持。

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.