Thermo-Finnigan Deca-XP LCMS for proteomics

描述（由申请人提供）：要求资金购买热-芬尼根液相色谱-质谱仪（LCMS），电喷雾电离（ESI），使用Deca XP四极离子阱分析仪，能够进行MS/MS碰撞诱导解离（MS2）和MSn碎片。所要求的系统是thermal - finnigan Proteome X配置，该配置捆绑了用于分离复杂肽混合物的二维色谱的HPLC设备，并包括用于控制HPLC的特定软件包，离子阱，包括数据依赖获取，以及促进蛋白质组学应用的特定软件。该软件包括关键数据库搜索算法（SEQUEST），用于根据这些蛋白质的消化肽的光谱识别混合物中的蛋白质。虽然有较旧的三扇形四极杆ESI-LCMS和基质辅助激光解吸电离飞行时间（MALDI-TOF），但这些仪器是离子阱ESI-LCMS的补充，不能执行该仪器所需的必要功能。只有当前一代离子阱ESI-LCMS离子阱技术具有灵敏度和速度，可以对复杂混合物中的肽进行必要的快速MS/MS光谱收集，从而通过序列识别，通过SEQUEST数据库搜索原蛋白，包括对这些蛋白的修饰。来自佛蒙特大学（UVM）不同部门和学院的各种研究人员，由几个不同的NIH研究所资助，聚集在一起要求以通用方式配置离子阱ESI-LCMS系统，但重点是解决蛋白质组学和蛋白质化学问题。在UVM或整个州都没有这样的工具可用。若干资助项目的调查已取得成功，现在需要回答需要所需工具的新的和更机械的问题。其他研究人员已经开始进行蛋白质组学研究，但这些举措受到外部合作和服务收费安排的限制。这些研究人员将大大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.