Evolutionary Origins of Protein Regulation

蛋白质调控的进化起源

• 批准号: 10443565
• 负责人: Brian Gene Miller
• 金额: $ 29.82万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10443565
• 关键词: Address; Allosteric Regulation; Amino Acid Sequence; Binding; Biochemical; Biological Models; Biology; Biophysics; Cell physiology; Data; Diabetes Mellitus; Disease; Enzyme Kinetics; Enzymes; Event; Evolution; Gene Duplication; Glucokinase; Goals; Growth; Health; Heterogeneity; Homeostasis; Human; Hyperinsulinism; Individual; Knowledge; Ligand Binding; Link; Mammals; Metabolic Diseases; Mission; Modernization; Molecular; Molecular Conformation; Mutation; Natural Selections; Peptidoglycan; Phenotype; Physiology; Play; Process; Protein Analysis; Protein Conformation; Protein Family; Proteins; Proteolysis; Public Health; Recycling; Regulation; Research; Resolution; Role; Sampling; Scaffolding Protein; Structure; Surface; System; Testing; United States National Institutes of Health; Variant; Vertebrates; Work; X-Ray Crystallography; biophysical analysis; blood glucose regulation; empowered; experimental study; genetic regulatory protein; glucokinase regulatory protein; glucose sensor; human disease; insight; novel; polypeptide; protein protein interaction; reconstruction; response; small molecule; stem

Protein regulation is ubiquitous in biology. It plays an essential role in controlling and coordinating a variety of cellular processes including replication, division and growth. Protein mis-regulation has been implicated in many human diseases. The emergence of new regulatory strategies often requires protein conformational heterogeneity. For example, allostery requires the ability to toggle between two (or more) distinct polypeptide structures. Similarly, a protein's ability to interact with multiple binding partners often requires sampling a range of conformations. Past experimental studies of individual polypeptides have yielded a molecular level understanding of many conformation-dependent regulatory processes. Despite this fact, we know little about how amino acid sequence changes serve to alter a protein's conformational landscape, such that new regulatory strategies can be realized. The goal of this proposal is to illuminate how evolutionary trajectories, shaped by natural selection, provide access to new protein conformations, which facilitate the emergence of novel regulatory mechanisms. As a model system, we will study the evolution of regulation within the glucokinase (GCK) protein family. GCK is regulated by several mechanisms that require access to multiple conformations, and GCK is evolutionarily tractable – its orthology is established and sequences from a range of species are available. We will use ancestral sequence reconstruction to resurrect ancestral versions of GCK and it's regulatory protein, GKRP. We will perform structural, biochemical and biophysical analyses of these proteins to illuminate the functional transitions that afford access to new protein conformations, which facilitate novel modes of regulation. Understanding the evolutionary origins of GCK's diverse regulatory processes will provide direct insight into the emergence of physiology and disease, as GCK is the primary glucose sensor in humans and a key determinant of glucose homeostasis in vertebrates. Fundamental questions that will be addressed in this proposal include: (1) How does a disordered polypeptide region evolve from an ordered precursor? (2) How do sequence changes expand the conformational landscape of a protein? (3) What events led to the emergence of allostery? (4) How does a new regulatory protein-protein interaction arise from two non-interacting ancestral proteins?

蛋白质调控在生物学中无处不在。它在控制和协调各种 包括复制、分裂和生长。蛋白质的错误调节 在许多人类疾病中。新的调控策略的出现往往需要蛋白质构象 异质性例如，变构需要能够在两个（或更多个）不同的 多肽结构类似地，蛋白质与多个结合伴侣相互作用的能力通常需要 对一系列构象进行采样过去对单个多肽的实验研究已经产生了 在分子水平上理解许多构象依赖的调控过程。尽管如此，我们 对于氨基酸序列的变化如何改变蛋白质的构象景观， 从而实现新的监管策略。本提案的目的是阐明 由自然选择塑造的进化轨迹提供了获得新蛋白质构象的途径， 促进新的调节机制的出现。作为一个模型系统，我们将研究 葡萄糖激酶（GCK）蛋白家族内的调节。GCK受几种机制调节， 需要获得多种构象，GCK在进化上是易于处理的-它的直系亲属是建立在 并且可获得来自一系列物种的序列。我们将使用祖先序列重建， 复活GCK及其调节蛋白GKRP的祖先版本。我们将进行结构， 这些蛋白质的生物化学和生物物理分析，以阐明功能转变， 获得新的蛋白质构象，促进新的调控模式。了解 GCK的多样性调节过程的进化起源将提供直接洞察的出现， 由于GCK是人体中的主要葡萄糖传感器和葡萄糖的关键决定因素， 脊椎动物体内平衡本提案将解决的基本问题包括：（1）如何 一个无序的多肽区域是从一个有序的前体进化而来的吗？(2)序列如何变化 扩展了蛋白质的构象(3)什么事件导致了变构的出现？（四） 一个新的调节蛋白质-蛋白质相互作用是如何从两个不相互作用的祖先蛋白质中产生的？