Activation and Regulation of Bacterial Tyrosine Kinases

细菌酪氨酸激酶的激活和调节

• 批准号: 1937937
• 负责人: Ranajeet Ghose
• 金额: $ 110.71万
• 依托单位: CUNY City College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1937937&HistoricalAwards=false
• 关键词: Activation; Regulation; Bacterial; Tyrosine; Kinases

Title: Activation and Regulation of Bacterial Tyrosine KinasesThe transfer of the terminal phosphate group from adenosine triphosphate (ATP) to specific tyrosine residues on defined protein targets has profound regulatory effects on all the processes that sustain life in higher eukaryotes. Indeed, there are nearly a hundred distinct human enzymes, known as tyrosine kinases, that facilitate this transfer. Dysregulation of tyrosine kinase activity has been implicated in a variety of human diseases including several cancers making these enzymes ideal targets for therapeutic intervention. Tyrosine phosphorylation driven signal transduction was considered to be restricted to the eukaryotic domain of life and indeed the presence of many distinct tyrosine kinases was believed to be the hallmark of multicellularity. However, recent discoveries have provided clear evidence that bacterial cells also encode numerous unique tyrosine kinases that play a similarly important role in bacterial physiology. The largest family of these bacterial tyrosine kinases, the BY-kinases, are conserved across the bacterial kingdom, contain none of the sequence signatures characteristic of eukaryotic tyrosine kinases, are structurally unique, and are activated and regulated through distinct, yet incompletely understood, mechanisms. The research proposed, using an integrated application of experimental and computational tools will decipher in terms of structure and mechanism, the activation and regulation of BY-kinase function. This research will provide multi-disciplinary training to a broad range of scholars at various career stages ranging from high-school students to postdoctoral trainees. The work will be facilitated by the outstanding intellectual environment at the City College of New York enabled by a student body that reflects the unique and diverse demographics of New York City.The long-term goal of the research is to understand the activation, activity and regulation of BY-kinases in atomic detail. Towards that goal, the mechanism/s employed by the catalytic subunit of BY-kinases in achieving efficient auto-phosphorylation and the nature of their interaction with counteracting PTPs to reverse this covalent modification, will be determined. The proposed research will utilize solution-state nuclear magnetic resonance and high-resolution mass-spectrometric techniques informed by all-atom computational studies and validated by biochemical experiments in vitro and in cell. The proposed studies will provide an expanded view of protein phosphorylation in life’s kingdoms through detailed insight into a divergent platform utilized by nature to perform this simple chemistry that has profound biological consequences. The archetypal BY kinase, Escherichia coli Wzc, and its cognate protein tyrosine phosphatase, Wzb will be utilized in these studies. Deploying proteins from a non-pathogenic, genetically tractable organism will allow the robust validation of the in vitro biophysical/biochemical insights within the cellular context. This project is supported by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division in the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

标题：细菌酪氨酸基纳氏菌的激活和调节末端磷酸基团从三磷酸腺苷（ATP）转移到特定的酪氨酸靶标，对在较高真核生物中维持寿命的所有过程都具有深远的调节作用。实际上，有将近一百个不同的人类酶，称为酪氨酸激酶，可以促进这种转移。酪氨酸激酶活性的失调已在多种人类疾病中实施，包括几种癌症，使这些酶成为热干预的理想靶标。酪氨酸磷酸化驱动的信号转导被认为仅限于生命的真核生物领域，实际上，人们认为许多不同的酪氨酸激酶被认为是多细胞性的标志。然而，最近的发现提供了明确的证据，表明细菌细胞还编码了许多独特的酪氨酸激酶，这些激酶在细菌生理学中起着类似重要的作用。这些细菌酪氨酸激酶的最大家族（副激酶）在整个细菌界都保守，不包含真核酪氨酸激酶的特征的序列特征，在结构上是独特的，并且通过独特的，但不完全理解的机制受到激活和调节。该研究提出的，使用实验和计算工具的综合应用将在结构和机制，激活和调节副酶功能方面解密。这项研究将为各个职业阶段的广泛学者提供多学科的培训，从高中生到博士后学员。这项工作将由纽约市城市学院的杰出知识环境准备，由一个反映纽约市独特而潜水员人口的学生团体，这项研究的长期目标是了解原子质细节的激活，活动和调节。为了实现这一目标，将确定将确定副激酶的催化亚基在实现有效的自磷酸化方面使用的机制，以及它们与抵抗PTP的相互作用的性质，以扭转这种价值修饰。拟议的研究将利用溶液状态核磁共振和全原子计算研究告知的高分辨率质谱技术，并通过体外和细胞中的生化实验验证。拟议的研究将通过详细洞悉自然界利用的不同平台来执行这种简单的化学反应，从而扩大生活王国蛋白质磷酸化的看法。这些研究将使用激酶，大肠杆菌WZC及其同源蛋白酪氨酸磷酸酶的原型。从非致病性，一般可拖延的组织中部署蛋白质将允许在细胞环境中对体外生物物理/生化洞察的良好验证。该项目得到了生物科学局的分子和细胞生物科学司的分子生物物理学簇的支持。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的审查标准来评估的。

项目成果

Long-range dynamic correlations regulate the catalytic activity of the bacterial tyrosine kinase Wzc

• DOI: 10.1126/sciadv.abd3718
• 发表时间: 2020-12-01
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Hajredini, Fatlum;Piserchio, Andrea;Ghose, Ranajeet
• 通讯作者: Ghose, Ranajeet