Molecular Profiling and Characterization

分子图谱和表征核心是UIUC细胞-细胞神经蛋白质组学中心的核心 发信号。了解大脑化学在健康、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。大的 化学信使的种类及其化学和物理性质的多样性需要一种复合体 一套测量技术和综合方法，用于解剖细胞到细胞的分子组成 蜂窝通讯。质谱学(MS)是精确的分子图谱和 描述复杂的神经系统。分子生物学和分子生物学的主要任务 表征的核心是使用基于MS的方法来增强我们对成瘾相关过程的理解 提供细胞-细胞间分子驱动因素的定性、定量和功能数据的分析 发信号。定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，该核心执行具有最高数据质量的已定义研究，并提供定制的MS 实现位于或接近当前技术极限的用户特定实验的方法。我们有 出色的分析能力和完善的专业知识，可有效探测各种信令 组织内的成分、单个细胞及其提取物和释放物具有空间和时间分辨率。 这些措施充分利用了专业人员精心的研究设计和严格的执行 在整个中心开发和实施经过验证的方法。伊利诺伊大学伊利诺伊分校 厄巴纳-香槟大学和西北大学，我们配备齐全的高级套房。 分辨率、串联质谱仪和采样能力，适用于成像和分子 分析大分子范围内的神经肽、代谢物、蛋白质和蛋白质复合体。 对于蛋白质组学，已定义的研究设计被归类为“自下而上”和“中下而下”的猎枪方法 在MS分析之前对蛋白质进行不同程度的消化，以及“自上而下”的方法 将完整的蛋白质或完整的蛋白质复合体引入到质谱仪中进行结构分析。 我们在以下背景下探讨了这些方法的互补性 中心支持的特定研究项目，以最大限度地提高鉴定的分子特异性和覆盖面 和蛋白质及其翻译后修饰的定量。未来的研究将受益于高 关于获得的生物活性神经肽和代谢物水平的相对和绝对量的分辨率信息 在目标或发现操作模式下。我们还使用了MS成像方法来照亮 “隐藏”的多肽组和代谢组，以阐明与药物滥用有关的关键分子和途径并提供信息 对相关疾病的治疗，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.