Molecular Profiling and Characterization

分子分析和表征核心是UIUC细胞神经蛋白质组学中心的核心 发信号了解脑化学在健康，疼痛管理，药物成瘾和戒断中的作用 需要了解大脑中的分子及其相关的神经化学途径。大 化学信使的分类及其化学和物理性质的多样性需要一个复杂的 一套测量技术和综合方法，用于解剖细胞到细胞的分子组分， 细胞通讯质谱（MS）是用于精确分子谱分析的核心技术， 复杂神经系统的特征。分子特征分析的主要使命和 表征核心是使用基于MS的 分析，以提供有关细胞间相互作用中所涉及的分子驱动因素的定性、定量和功能数据。 信号位于采样和分离核心与生物信息学，数据分析和 预测建模核心，该核心以最高的数据质量执行定义的研究，并提供定制的MS 用于实现处于或接近当前技术极限的用户特定实验的方法。我们有 出色的分析能力和有据可查的专业知识，可有效探测各种信号 组织内的组分、单个细胞及其提取物和释放物具有空间和时间分辨率。 这些测量利用了精心的研究设计和专业人员的严格执行， 开发和实施经过验证的方法。伊利诺伊大学 厄巴纳-香槟和西北大学，我们配备了一个广泛的套房高- 分辨率、串联质谱仪和采样能力，适用于成像和分子 神经肽、代谢物、蛋白质和蛋白质复合物的分析，涵盖广泛的分子量范围。 对于蛋白质组学，定义的研究设计分为“自下而上”和“中间向下”鸟枪法 其中蛋白质在MS分析之前被消化到不同程度，以及“自上而下”方法，其中 将完整蛋白质或完整蛋白质复合物引入质谱仪中进行结构分析， 我们利用这些方法的互补性的背景下， 特定中心支持的研究项目，以最大限度地提高分子特异性和识别覆盖率 和蛋白质及其翻译后修饰的定量。未来的研究将受益于高- 获得的生物活性神经肽和代谢物水平的相对量和绝对量的分离度信息 在目标操作模式或发现操作模式中。我们还采用MS成像方法来阐明 “隐藏”的肽组和代谢组，以阐明药物滥用和信息的关键分子和途径 治疗相关疾病，如药物成瘾和戒断。

The Molecular Profiling and Characterization Core is the heart of the UIUC Neuroproteomics Center on Cell-Cell Signaling. Understanding the role of brain chemistry in health, pain management, drug addiction, and withdrawal requires knowledge of the molecules within the brain and their associated neurochemical pathways. The large assortment of chemical messengers and the diversity of their chemical and physical properties require a complex suite of measurement techniques and integrated approaches for dissecting the molecular components of cell-to- cell communication. Mass spectrometry (MS) is a central technique for precise molecular profiling and characterization of complex neurological systems. The primary mission of the Molecular Profiling and Characterization Core is to augment our understanding of addiction-associated processes using MS-based analytics to provide qualitative, quantitative, and functional data on the molecular drivers involved in cell-cell signaling. Positioned between the Sampling and Separation Core and Bioinformatics, Data Analytics and Predictive Modeling Core, this core executes defined studies with maximal data quality and provides tailored MS methods for enabling user-specific experiments that lie at or near current technological limits. We have the outstanding analytical capabilities and well-documented expertise to effectively probe diverse signaling components within tissues, individual cells, and their extracts and releasates with spatial and temporal resolution. These measurements capitalize on the careful study design and rigorous execution by professional staff capable of developing and implementing validated methods across the Center. Between the University of Illinois at Urbana-Champaign and Northwestern University, we are well-equipped with an extensive suite of high- resolution, tandem mass spectrometers and sampling capabilities that are suitable for imaging and molecular analysis of neuropeptides, metabolites, proteins, and protein complexes across a wide molecular-weight range. For proteomics, defined study designs are categorized as “Bottom-Up” and “Middle-Down” shotgun approaches in which proteins are digested to various extents prior to MS analysis, and the “Top-Down” approach in which the intact proteins or whole protein complexes are introduced into the mass spectrometer for structural characterization by tandem MS. We exploit the complementarity of these approaches within the context of specific Center-supported research projects to maximize the molecular specificity and coverage for identification and quantitation of proteins and their post-translational modifications. Future studies will benefit from high- resolution information on relative and absolute amounts of bioactive neuropeptide and metabolite levels obtained in either targeted or discovery modes of operation. We also employ MS imaging approaches to illuminate the “hidden” peptidome and metabolome to elucidate key molecules and pathways involved in drug abuse and inform treatments for associated conditions such as drug addiction and withdrawal.