Thermo-Finnigan Deca-XP LCMS for proteomics

描述(由申请人提供)：申请资金购买一台电喷雾电离(ESI)的Thermo-Finnigan液相色谱质谱计(LCMS)，该仪器使用Deca XP四极离子陷阱分析仪，能够执行MS/MS碰撞诱导解离(MS2)和MSN碎裂。所要求的系统是Thermo-Finnigan Proteome X配置，它捆绑了将执行二维(2D)层析以分离复杂多肽混合物的高效液相色谱设备，并包括专门用于控制高效液相色谱的软件包、离子陷阱，包括数据依赖采集，以及专门用于促进蛋白质组学应用的软件。该软件包括关键数据库搜索算法(SEQUEST)，用于基于这些蛋白质消化中的多肽的光谱来识别混合物中的蛋白质。虽然有较旧的三扇区四极杆电离-飞行时间质谱仪和基质辅助激光解吸电离飞行时间(MALDI-TOF)，但这些仪器是对所要求的离子陷阱电离-飞行时间质谱仪的补充，不能执行需要该仪器的必要功能。只有当前一代的离子陷阱ESI-LCMS离子陷阱技术具有对复杂混合物中的多肽进行必要的快速MS/MS光谱收集以根据其序列识别SEQUEST数据库搜索原始蛋白质，包括对这些蛋白质的修饰所需的灵敏度和速度。来自佛蒙特大学(UVM)不同系和学院的不同研究人员，在几个不同的NIH研究所的资助下，聚集在一起，要求以一种多功能的方式配置这种离子陷阱ESI-LCMS系统，但专注于解决蛋白质组和蛋白质化学问题。在密歇根州立大学或整个州都没有这种仪器。几个受资助的项目在其调查中取得了成功，现在需要回答新的和更机械化的问题，这些问题需要所要求的工具。其他研究人员已经开始了蛋白质组学研究，但这些倡议受到外部合作和收费服务安排的限制。如果这些研究人员在内部拥有必要的仪器设备，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，这些研究人员将大大受益。

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.