Protein Phosphatase 1 Holoenzyme Formation

蛋白磷酸酶 1 全酶形成

基本信息

批准号：
10671729
负责人：
Wolfgang Peti
金额：
$ 38.72万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
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：
2022-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10671729
关键词：
ATP phosphohydrolase Area Binding Biochemical Biogenesis Biological Biological Process Biology Biophysics Cells Cellular biology Complex Cysteine Data Development Disease Dissociation Drug Targeting Enzymes Essential Genes Eukaryota Eukaryotic Cell Excision Genetic Goals Health Holoenzymes Hydrolysis Individual Laboratories Location Malignant Neoplasms Metals Mitosis Molecular Molecular Chaperones Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Post-Translational Protein Processing Protein Deficiency Protein Dephosphorylation Protein Inhibition Protein phosphatase Proteins Reaction Regulation Research Research Personnel Research Project Grants Role Serine/Threonine Phosphorylation Signal Transduction Structure Substrate Specificity Testing Threonine Phosphorylation Site Work Yeasts biophysical tools cell growth dimer experimental study human disease inhibitor inorganic phosphate insight novel novel strategies protein phosphatase inhibitor-1 response structural biology tool ubiquitin-protein ligase yeast genetics

ATP phosphohydrolase Area Binding Biochemical Biogenesis Biological Biological Process Biology Biophysics Cells Cellular biology Complex Cysteine Data Development Disease Dissociation Drug Targeting Enzymes Essential Genes Eukaryota Eukaryotic Cell Excision Genetic Goals Health Holoenzymes Hydrolysis Individual Laboratories Location Malignant Neoplasms Metals Mitosis Molecular Molecular Chaperones Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Post-Translational Protein Processing Protein Deficiency Protein Dephosphorylation Protein Inhibition Protein phosphatase Proteins Reaction Regulation Research Research Personnel Research Project Grants Role Serine/Threonine Phosphorylation Signal Transduction Structure Substrate Specificity Testing Threonine Phosphorylation Site Work Yeasts biophysical tools cell growth dimer experimental study human disease inhibitor inorganic phosphate insight novel novel strategies protein phosphatase inhibitor-1 response structural biology tool ubiquitin-protein ligase yeast genetics

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项目摘要

ABSTRACT Phosphorylation is one of the most ubiquitous, reversible posttranslational modifications in cells. The enzymes responsible for controlling the phosphorylation state of the cell are kinases, which catalyze the transfer of the γ-phosphate moiety of ATP to substrates, and phosphatases, which catalyze the reverse hydrolysis reaction, the removal of the phosphate moiety from phosphorylated substrates. Thus, phosphatases dynamically reverse the effects of kinases. Because phosphorylation is critical for all biological processes from cell growth to differentiation to development, the location and duration of the reciprocal actions of kinases and phosphatases must be exquisitely regulated both temporally and spatially within the cell. Consequently, when this tight regulation is disrupted, dysregulation of phosphorylation signaling ensues and the consequence is most often disease. Deletion of either one of two PP1 regulators—SDS22 (PPP1R7) or Inhibitor-3 (I3; PPP1R11 or Ypi1 in yeast)—is lethal in yeast (essential genes), highlighting their biological significance. However, since their discovery, different biological roles have been assigned to SDS22 and I3, including roles in mitosis (SDS22), E3 ligase functionality (I3), PP1 biogenesis, among others. Thus, while it is clear that SDS22 and I3 are essential PP1 regulators, their true biological function(s) and especially their mechanism(s) of action are still unknown. This has hindered progress in understanding their roles in PP1 biology. In cells, these proteins form both heterodimeric (SDS22:PP1 and I3:PP1) and a heterotrimeric (SDS22:I3:PP1; SIP) PP1 complex. The structure and function(s) of the individual dimeric complexes, if and how the structure and function(s) of the trimeric complex differs from those of the dimeric complexes and the role(s) of each complex in PP1 holoenzyme formation are major questions in the field. Further, additional data suggest that dissociation of the SIP complex requires the AAA+ ATPase p37/p97. However, the molecular details of SIP complex dissociation have also remained elusive. The presented research project uses a powerful integrated approach that combines structural biology with biochemical and cell biology experiments to obtain novel insights into the molecular mechanisms used by these regulators to control PP1 activity and direct PP1 holoenzyme assembly. Because PP1 holoenzymes have critical roles in human diseases, the proposed work will provide novel strategies for selectively inhibiting PP1 activity by targeting the PP1 holoenzyme formation and subunit exchange, which is essential for understanding how distinct PPPs contribute to disease.

抽象的磷酸化是细胞中最普遍，可逆的翻译后修饰之一。酶负责控制细胞的磷酸化状态的是激酶，这会催化 γ-磷酸ATP的磷酸部分和底物和磷酸酶，这些磷酸酶催化反向水解反应，从磷酸化的底物中去除磷酸盐部分。那，磷酸酶动态逆转激酶的作用。因为磷酸化对于从细胞生长到所有生物过程至关重要分化与发育，激酶和磷酸酶相互作用的位置和持续时间必须在细胞内临时和空间上的临时调节。因此，当这个紧绷调节受到破坏，随之而来的磷酸化信号失调，最常见的结果是疾病。删除两个PP1调节器之一-SDS22（PPP1R7）或抑制剂3（i3; PPP1R11或YPI1）在酵母中） - 在酵母中致命（基本基因），突出了它们的生物学意义。但是，由于他们发现，已将不同的生物学作用分配给SDS22和i3，包括有丝分裂的作用（SDS22）， E3连接酶功能（I3），PP1生物发生等。那很明显SDS22和i3是必需的PP1调节剂，其真正的生物学功能，尤其是其作用机制仍然是未知。这阻碍了理解他们在PP1生物学中的作用的进展。在细胞中，这些蛋白质形成杂二聚体（SDS22：PP1和I3：PP1）和异三聚体（SDS22：i3：pp1; sip）PP1复合物。这单个二聚体复合物的结构和功能，如果以及如何以及如何的结构和功能三聚体复合物与二聚体复合物的差异和pp1中每个复合物的作用全酶形成是该领域的主要问题。此外，其他数据表明 SIP复合物需要AAA+ ATPase p37/p97。但是，SIP复合物分离的分子细节也仍然难以捉摸。提出的研究项目使用了强大的综合方法将结构生物学与生化和细胞生物学实验相结合，以获得对该的新见解这些调节剂使用的分子机制来控制PP1活性和直接PP1全酶组件。由于PP1全酶在人类疾病中具有关键作用，因此拟议的工作将提供新颖通过靶向PP1全酶形成和亚基来选择性抑制PP1活性的策略交换，这对于了解不同的PPP对疾病的贡献至关重要。