Thermo-Finnigan Deca-XP LCMS for proteomics

描述（由申请人提供）：需要资金购买配备电喷雾电离 (ESI) 功能的 Thermo-Finnigan 液相色谱-质谱仪 (LCMS)，使用 Deca XP 四极杆离子阱分析仪，能够执行 MS/MS 碰撞诱导解离 (MS2) 和 MSn 碎片。所需的系统是 Thermo-Finnigan Proteome X 配置，该配置捆绑了 HPLC 设备，该设备将执行二维 (2D) 色谱法以分离复杂的肽混合物，并包括专用于控制 HPLC 的软件包、离子阱（包括数据依赖性采集）和专用于促进蛋白质组学应用的软件。该软件包括关键的数据库搜索算法 (SEQUEST)，用于根据这些蛋白质消化物中肽的光谱来识别混合物中的蛋白质。尽管有较旧的三扇区四极杆 ESI-LCMS 和基质辅助激光解吸电离飞行时间 (MALDI-TOF) 可用，但这些仪器是对所需离子阱 ESI-LCMS 的补充，无法执行该仪器所需的必要功能。只有当前一代离子阱 ESI-LCMS 离子阱技术具有足够的灵敏度和速度，能够对复杂混合物中的肽进行必要的快速 MS/MS 谱图收集，从而通过序列识别 SEQUEST 数据库搜索原始蛋白质，包括对这些蛋白质的修饰。在几个不同的 NIH 研究所的资助下，来自佛蒙特大学 (UVM) 不同院系和学院的众多研究人员齐聚一堂，要求以多功能方式配置离子阱 ESI-LCMS 系统，但重点是解决蛋白质组和蛋白质化学问题。 UVM 或整个州都没有这样的仪器。一些受资助的项目在调查中已经取得了成功，现在需要回答新的、更机械的问题，而这些问题需要所要求的工具。其他研究人员已经开始蛋白质组学研究，但这些举措受到外部合作和按服务收费安排的限制。这些研究人员将受益于拥有必要的内部仪器，他们、他们的同事和学生可以直接使用和学习他们的研究越来越依赖的技术。

DESCRIPTION (provided by applicant): Funds are requested to purchase a Thermo-Finnigan liquid chromatograph-mass spectrometer (LCMS) with electrospray ionization (ESI) using a Deca XP quadrupole ion trap analyzer capable of performing MS/MS collision induced disassociation (MS2) and MSn fragmentation. The requested system is the Thermo-Finnigan Proteome X configuration that bundles HPLC equipment that will perform 2-dimensional (2D) chromatography for separating complex peptide mixtures and includes software packages specific for controlling the HPLC, the ion trap, including data dependent acquisition, and software specific to facilitate proteomics applications.The software includes critical database search algorithms (SEQUEST) for identifying proteins in mixtures on the basis of the spectra of peptides in the digest of these proteins. Although there are older triple sector quadrupole ESI-LCMS and a matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) are available, these instruments are complementary to the requested ion trap ESI-LCMS and cannot perform the necessary functions for which this instrument is required. Only the current generation ion trap ESI-LCMS ion trap technology has the sensitivity and speed to perform the necessary rapid MS/MS spectra collection of peptides in complex mixtures to identify by their sequences the SEQUEST database search the originating proteins, including modifications to these proteins. A variety of investigators from different departments and colleges at the University of Vermont (UVM), funded by several different NIH institutes, have come together to request this ion trap ESI-LCMS system configured in a versatile manner, but focused to solve proteomic and protein chemistry problems. No such instrument is available either at UVM or in the entire state. Several of the funded projects have reached successful points in their investigations that now beg new and more mechanistic questions be answered that require the requested instrument. Other investigators have already begun proteomic studies, but these initiatives are limited by constraints of external collaboration and fee-for-service arrangements. These investigators would greatly benefit from having the necessary instrumentation in-house where they, their colleagues and students could directly use and learn the techniques for which their research is becoming increasingly more dependent.