Focal adhesion protein analysis from designed surfaces

描述(由申请人提供)：识别负责细胞黏附、突起和回缩的蛋白质对于理解细胞迁移过程至关重要。质谱学是一种非常适合检测蛋白质及其翻译后修饰的分析工具。然而，大多数基于质谱学的方法来研究这些成分依赖于溶液相操作，即使许多负责介导细胞迁移的蛋白质并不作为细胞质中的可溶性实体发挥作用。相反，细胞的内膜为蛋白质组装成称为焦点黏附复合体的结构提供了一个独特的环境。细胞迁移是由连接细胞与其所在表面的粘附物的类型、数量和接触强度来调节的。因此，表面是研究这一过程的一种更具生理学相关性的媒介。本文提出的研究将描述在自组装单分子膜上重组的焦点黏附复合体的新方法。这些表面将被设计成呈现电活性运动，可用于特定地释放分析物，并便于通过质谱学进行检测。最后，将讨论这些方法在假设驱动下的应用。

DESCRIPTION (provided by applicant): The identification of proteins responsible for cell adhesion, protrusion, and retraction is critical to understanding the process of cell migration. Mass spectrometry is an analytical tool well-suited for detection of proteins and their post-translation modifications. Nevertheless, the majority of mass spectrometry-based methods to study these components rely on solution phase manipulations, even though many of the proteins responsible for mediating cell migration do not function as soluble entities in the cytoplasm. Rather, the inner membrane of a cell provides a unique environment for proteins to assemble into structures called focal adhesion complexes. Cell migration is mediated by the type, number, and contact strength of the adhesions connecting the cell to the surface on which it resides. Therefore, surfaces are a more physiologically relevant medium to study this process. The research proposed here will describe new methods to characterize focal adhesion complexes reconstituted on self-assembled monolayers. These surfaces will be designed to present electroactive moeities that can be used to specifically release the analyte and facilitate detection by mass spectrometry. Finally, hypothesis-driven applications of these methods will be discussed.