A New Pradigm for the Rational Expression of Integral Membrane Proteins

摘要 整体膜蛋白(IMP)具有重要的科学和医学意义，因为它们支配着 营养物质和信息在细胞膜上的流动，因为它们构成了最大类别的治疗药物 目标。然而，IMPS的结构和生物物理特性远远落后于其他蛋白质类别 无法可靠地生产大量用于实验研究的IMP。中国的根本差距 目前还不存在理解氨基酸和核苷酸之间联系的框架 IMP及其表达水平的序列，技术上的根本差距是 没有成功的战略存在 用于一般预测和改善IMP的表达水平。 目前的工作将消除这些根本差距 在知识和技术方面，通过(I)实验方法来表征IMP的表达 跨多个蛋白质家族和表达系统的水平；(Ii)严格的数据驱动策略 不同的IMP序列属性对表达的作用，以及(Iii)粗粒度分子动力学方法使 对作为IMP表达瓶颈的关键机械步骤的模拟--基于无偏见的生物时间尺度。 加州理工学院米勒和克莱蒙斯实验室提议的合作具有得天独厚的优势，可以在这方面取得成功 努力，为小鬼理性表达的新范式创造机会。拟议研究的成功 将通过为增加IMP表达提供强大且可访问的计算工具来产生转型影响 对于研究界来说，通过推进这一大类和重要的蛋白质的基本特征， 并通过加快医疗和制药应用。

ABSTRACT Integral membrane proteins (IMPs) are of both fundamental scientific and medical importance, because they govern the flow of nutrients and information across cell membranes and because they comprise the largest class of therapeutic drug targets. However, the structural and biophysical characterization of IMPs lags far behind that of other protein classes, due to the inability to reliably produce significant quantities of IMPs for experimental studies. The fundamental gap in knowledge is that no framework currently exists for understanding the connection between the amino-acid and nucleotide sequence of an IMP and its expression level, and the fundamental gap in technology is that no successful strategies exist for the general prediction and improvement IMP expression levels. The current work will remove these fundamental gaps in knowledge and technology via the innovative combination of (i) experimental methods to characterize IMP expression levels, spanning across multiple protein families and expression systems, (ii) rigorous data-driven strategies to disentangle the role of diverse IMP sequence properties on expression, and (iii) coarse-grained molecular dynamics methods to enable the simulation - on unbiased biological timescales - of key mechanistic steps that act as bottlenecks to IMP expression. The proposed collaboration of the Miller and Clemons laboratories at Caltech is uniquely positioned to succeed in this effort, creating the opportunity for a new paradigm for the rational expression of IMPs. Success of the proposed research will have transformational impact by providing powerful and accessible computational tools for increased IMP expression to the research community, by advancing the fundamental characterization of this large and important class of proteins, and by accelerating medical and pharmaceutical applications.