Evolution of new protein function in the multi-protein, multi-functional Toll-like receptor 4 complex

多蛋白、多功能Toll样受体4复合物中新蛋白功能的进化

基本信息

批准号：
10625469
负责人：
Michael Jonathan Harms
金额：
$ 36.49万
依托单位：
UNIVERSITY OF OREGON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10625469
关键词：
Acylation Address Amphibia Animal Model Animals Binding Binding Sites Biochemical Biochemistry Bioinformatics Biological Models Biology Biophysics CD14 gene Complex Conflict (Psychology)Depressed mood Disease Drug Targeting Evolution Exhibits Exposure to Genetic Goals Health Homologous Gene Human Human Biology Immune Infection Inflammation Inflammatory Investigation Lipopolysaccharides Mammals Measurement Microbe Modernization Multiprotein Complexes Mutation Natural Immunity Osteichthyes Outcome Pattern Phylogenetic Analysis Play Process Proteins Role S100A9 gene Sampling Shapes Signal Transduction Site Specificity Sterility Structure Study models TLR4 gene Testing Tetrapoda Time Tissues Transferrin Vertebrates Work Zebrafish antagonist biophysical analysis experimental study fascinate in vivo insight interest lens member microbial mutation screening preservation pressure protein complex protein function reconstruction simulation

项目摘要

SUMMARY Understanding the evolution of protein function is a central goal in evolutionary biochemistry. Most mechanistic work on this problem has focused on single proteins with single functions; however, proteins often work as members of multi-functional, multi-protein complexes. How can evolution optimize one protein function without breaking another? How are new proteins integrated into multi-protein complexes? Answering these and other questions requires mechanistic studies of the natural evolution of multi-functional, multi-protein complexes. To answer these questions, we propose studying the evolution of Toll-like receptor 4 (TLR4)—a multi-protein, multi-functional protein complex found in vertebrate animals. This complex is central to how animals respond to both infection and tissue damage. It is the subject of intense interest, both from the perspective a basic biology and as a drug target for inflammatory disorders. The complex evolved in serial over hundreds of millions of years. In this work, we will pose—and answer—three questions: 1) How did the ability of the complex to recognize pro- inflammatory signals change over the last 300 million years? 2) What were the evolutionary forces and processes that led to these functional changes? 3) How did a completely new protein evolve in the ancestor of land animals and become integrated into the complex? To answer these questions, we will employ a combination of phylogenetic analysis, ancestral sequence reconstruction, high-throughput protein characterization, biochemical/biophysical studies, careful functional characterization, and in vivo experiments in zebrafish. This work will provide unprecedented understanding of the mechanisms—both biochemical and evolutionary— that lead to new function in multi-functional protein complexes. By specifically studying the evolutionary process that led to the human TLR4 complex, this work will also provide a lens through which we can interpret animal model studies of innate immunity and apply their findings to human biology. And, finally, an evolutionary approach is a powerful means to dissect how proteins work. Our results will provide deep insight into how this important protein complex functions and can be manipulated to achieve better human health outcomes.

概括了解蛋白质功能的进化是进化生物化学的核心目标。对这个问题的研究主要集中在具有单一功能的单一蛋白质上；然而，蛋白质通常作为多功能、多蛋白质复合物的成员，进化如何优化一种蛋白质功能。新蛋白质如何整合到多蛋白质复合物中？其他问题需要对多功能、多蛋白质的自然进化进行机制研究复合物。为了回答这些问题，我们建议研究 Toll 样受体 4 (TLR4) 的进化——一种多蛋白，在脊椎动物中发现的多功能蛋白质复合物，该复合物对于动物的反应至关重要。从基础生物学的角度来看，它是人们强烈关注的主题。作为炎症性疾病的药物靶点，该复合物经过数亿年的连续进化。在这项工作中，我们将提出并回答三个问题：1）复合体识别亲缘关系的能力如何？ 2）进化的力量和过程是什么？ 3）陆地动物的祖先中如何进化出一种全新的蛋白质？为了回答这些问题，我们将采用以下组合：系统发育分析、祖先序列重建、高通量蛋白质表征、生物化学/生物物理研究、仔细的功能表征以及斑马鱼的体内实验。这项工作将为人们提供前所未有的理解——无论是生化机制还是进化机制—— 通过专门研究进化，从而产生多功能蛋白质复合物的新功能。导致人类 TLR4 复合物的过程，这项工作还将提供一个镜头，通过它我们可以解释先天免疫的动物模型研究并将其研究结果应用于人类生物学。进化方法是剖析蛋白质工作原理的有力手段，我们的结果将提供深入的见解。研究这种重要的蛋白质复合物如何发挥作用并如何操纵以实现更好的人类健康结果。