ACQUISITION OF BRUKER DALTONICS REFLEX III MALDI TOF

该申请的具体目的是获得宾夕法尼亚大学最先进的高性能MALDI-TOF质谱仪，以满足研究人员对蛋白质、肽和其他生物分子研究的需求。具体来说，该提案的目的是扩大医学院现有蛋白质化学实验室（PCL）的能力。核心设施是最先进的微化学核心设施的后代，对整个大学来说，以下服务：1)肽合成，纯化和免疫原偶联2)肽修饰3)蛋白质纯化4)色氨酸制备5)蛋白质微测序6)质谱和7)蛋白质纯化和分析培训。该仪器将使我们能够a)获得低摩尔到飞摩尔多肽和蛋白质的高度精确的分子量数据，并允许在不断扩大的数据库中存在dna衍生序列信息的蛋白质的快速大规模定位鉴定。这种方法对于像PCL这样的全方位蛋白质核心实验室中可用的蛋白质测序资源的管理至关重要。后源衰减（PSD）和碰撞诱导解离（CID）特征与精确的质量数据相结合，即使存在于N端阻断的混合物中，也将提供肽的部分序列信息。蛋白质和其他生物分子的结构问题难以或无法用传统技术解决。值得注意的是，该仪器将集成到PCL进行的蛋白质化学的各个方面，如Edman测序和肽合成项目。总之，新仪器将为涉及天然产物和表达产物分析、蛋白质设计、肽合成、氨基酸测序、配体鉴定、翻译后修饰鉴定等问题提供高水平的置信度答案。，以及通过精确的质量定位、表征非蛋白生物分子来鉴定蛋白质。我们已经确定了10个主要用户研究项目的核心，这些项目来自宾夕法尼亚大学四所学院（艺术与科学、牙科医学、兽医医学和医学）。感兴趣的领域包括蛋白质-蛋白质相互作用，免疫系统的系统发育，基因表达和蛋白质设计，麻醉相关的蛋白质化学修饰，二硫键测定，以及细胞生长和分化的生物学。一些人已经从与我们核心的互动中受益，新的仪器将提高与各个参与实验室未来研究相关的结果的质量和量化。

The specific aim of this application is to acquire a state-of-the-art high performance MALDI-TOF mass spectrometer at the University of Pennsylvania to meet the needs of researchers concerned with studies of proteins, peptides, and other biomolecules. Specifically, the proposal intent is to expand the capabilities of the existing Protein Chemistry Laboratory (PCL) in the School of Medicine. The core facility is a state of art microchemical core facility offspring, to the entire University, the following services: 1) peptide synthesis, purification, and immunogenic coupling 2) peptide modifications 3) protein purification 4) tryptic peptide preparations 5) protein microsequencing 6.) mass spectroscopy and 7) training in protein purification and analysis. This instrument will enable us to a) obtain highly accurate molecular weight data on low pmole to femtomole amounts of peptides and proteins and permit rapid mass mapping identification of proteins whose cDNA-derived sequence information exists in the ever expanding databases. This approach is becoming essential to the management of protein sequencing resources available in an all-encompassing protein core laboratory like the PCL. Post source decay (PSD) and Collisionally induced dissociation (CID) features coupled with accurate mass data will provide partial sequence information on peptides even if present in a mixtures of N- terminally blocked. Structural problems on proteins and other biomolecules difficult or not approachable with conventional techniques will be addressable. Significantly the instrument will be integrated into all aspects of protein chemistries performed at the PCL such as Edman sequencing and peptide synthesis projects. In summary the new instrumentation will provide high-level-of-confidence answers for problems involving, natural product and expression product analysis, protein design, peptide synthesis, amino acid sequencing, ligand identification, post-translational modification identification., and protein identification through accurate mass mapping, characterization of non-protein biomolecules. We have identified a core of 10 major user research projects, from departments spanning four Penn schools (Art & Sciences, Dental Medicine, Veterinary Medicine and Medicine). The areas of interest include, protein-protein interactions, phylogeny of immune system, gene expression and protein design, anesthesia-related chemical modification of proteins, disulfide linkage determination, and biology of cell-growth and differentiation. Several people have already benefitted from interaction with our core and the new instrumentation will enhance the quality and quantify of results connected with future investigations of the various participating laboratories.