CAREER: Leveraging a fast-evolving kinase family to gain fundamental understanding of kinase evolution

激酶是细胞的关键组成部分，通过修饰蛋白质来发挥作用。这一过程是微调和调节的，因为激酶活性的变化可能是致命的，或者导致各种疾病。这意味着，像它们这样的信号分子往往进化得非常慢，因为它们必须既保持执行化学作用的能力，又要识别正确的目标。正因为如此，人们一直很难研究激酶及其信号网络。这个项目的研究目标是从寄生虫中开发出一个快速进化的激酶大家族，作为一个实验系统，可以与更典型的动物模型系统进行比较。这将使我们能够更好地定义管理激酶功能元件的生化规则。该项目的教育目标将(1)开发一门新课程，利用生物信息学和进化论的概念向本科生传授科学批判性思维技能；(2)调整这些课程材料，培训当地学校的高中教师，这些学校招收传统上在理科方面代表性不足的学生进入工程学。向高中生介绍基于探究的科学的目标教育目标将使本科生和高中生能够使用生物信息学工具提出并快速回答他们自己设计的问题。这将训练他们批判性思维，而批判性思维是科学探究的基础。这种批判性思维技能是现代教育的基本组成部分，并将在学生成长为公民的过程中进一步增强他们的能力。与正常的细胞内信号通路不同，弓形虫等球虫的效应器激酶的进化速度令正常酶相形见绌。这些球虫蛋白激酶进化很快，但作用于研究得很好的脊椎动物信号网络。我们将利用这个系统来发现新的生物化学和支配其进化的力量。这项研究将(1)确定一个球虫蛋白激酶家族在缺失甘氨酸环(Gly-loop)的情况下仍能够保持其催化活性的生化和结构机制，(2)利用祖先基因序列的重建来定义残基，这些残基使需要ATP稳定的活性蛋白激酶进化成不需要核苷酸的催化失活的假蛋白激酶，以及(3)确定一组直系球虫蛋白激酶如何改变其特异性，以靶向完全不同的脊椎动物途径。

Kinases are critical components of cells that work by modifying proteins. This process is finely tuned and regulated, as changes in kinase activity can be lethal or lead to a variety of diseases. This means that signaling molecules like them tend to evolve very slowly, because they must maintain both the ability to perform the chemistry as well as identify the right target. For this reason, it has been difficult to study kinases and their signaling networks. The research goals of this project are based on developing a large family of fast-evolving kinases from a parasite as an experimental system that can be compared to more typical animal model systems. This will allow us to better define the biochemical rules that govern the functional elements of kinases. This project's educational goals will (1) develop a new course to use bioinformatics and evolutionary concepts to teach undergraduates scientific critical thinking skills and (2) adapt these course materials to train high school teachers from local schools that enroll students who traditionally are underrepresented in science in engineering. The goal of introducing high school students to inquiry-based science The educational goals will empower students, both undergraduate and high school, to use bioinformatic tools to ask and quickly answer questions of their own devising. This will train them in the critical thinking that is the foundation of scientific inquiry. Such critical thinking skills are a fundamental part of modern education, and will further empower the students as they grow as citizensAs opposed to normal intracellular signaling kinases, effector kinases of the coccidian parasites such Toxoplasma gondii evolve at rates that dwarf normal enzymes. These coccidian kinases evolve quickly, but act on well-studied vertebrate signaling networks. We will harness this system to discover new biochemistry and the forces that govern its evolution. This research will (1) determine the biochemical and structural mechanism by which a family of coccidian kinases is able to retain its catalytic activity in spite of missing a Gly-loop, which is a motif that has been presumed to be absolutely required for kinase activity, (2) use the reconstruction of ancestral gene sequences to define the residues that enable active kinases that require ATP for stability to evolve into catalytically inactive pseudokinases that are stable without nucleotide, and (3) determine how sets of orthologous coccidian kinases have changed their specificity to target completely different vertebrate pathways.