Mapping the role of cofactors in the conformation of proteins using spectroscopy coupled with ion mobility mass spectrometry -summary to follow

我们获得了与Waters公司合作的资金，以适应市售的Synapt G2S HD质谱（BBSRC BB/L002655/1）。该仪器的核心是离子迁移细胞，在该仪器中可以进行imm -MS测量，就像任何Synapt质谱仪一样，但独特的是，我们能够捕获已选择的离子，以允许光学测量所谓的“光SRIG”（堆叠环离子指南）。这种光学测量可以采取几种形式，包括红外光谱，紫外/可见光谱，这将提供辅助因子和辅酶的结构分析，光解离蛋白质测序以及荧光探测整体构象。分子力学计算将产生与实验确定的结构相比较的结构。这个学生将使用这个新仪器来检测辅助因子在蛋白质构象中的作用，通过单独测量辅助因子，载脂蛋白和蛋白质的全息形式。学生将接受生物质谱、光学方法和分子力学方面的培训。他们将借调到沃特斯开发软件，以最好地解释该仪器的数据。这个尖端的跨学科科学项目是世界领先的，将培养学生在学术界和工业界都非常需要的一套技能。

We obtained funding to adapt a commercially available Synapt G2S HD MS (BBSRC BB/L002655/1) in collaboration with Waters Corp. The heart of this instrument is the ion mobility cell, IM-MS measurements are possible in this instrument, as for any Synapt mass spectrometer, but uniquely we are able to trap ions that have been conformer selected, to allow optical measurements in a so called 'photo SRIG' (stacked ring ion guide). Such optical measurements can take several forms, including IR-spectroscopy, UV/Vis-spectroscopy, which will provide structural analysis of co-factors and coenzymes, photo-dissociation to sequence the proteins as well as fluorescence to probe global conformations. Molecular mechanics calculations will generate structures to compare with those determined experimentally. This student will use this novel instrument to examine the role of cofactors on the conformations of proteins, by making measurements on the cofactor alone, the apo and the holo form of the protein. The student will be trained in biological mass spectrometry, in optical methods and in molecular mechanics. They will spend a secondment at Waters developing software to best interpret the data from this instrument. This cutting edge interdisciplinary science program is world leading and will train this student with a set of skills highly desirable both in academia and in industry.