Structural basis of PP2A phosphatase diseases

PP2A磷酸酶疾病的结构基础

基本信息

批准号：
10736835
负责人：
Yongna Xing
金额：
$ 36.2万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10736835
关键词：
Active Sites Adenine Affect Amino Acid Motifs Binding Biochemical Brain CREB1 gene Cancer Patient Catalytic Domain Cell physiology Cellular biology Common Core Complex Cryoelectron Microscopy Cyclic AMP Cyclic AMP-Dependent Protein Kinases Data Diagnosis Disease Dissection Event Family Feedback Forskolin Heart Diseases Holoenzymes Human Intellectual functioning disability Jordan Knowledge Link MAP Kinase Gene Malignant Neoplasms Missense Mutation Molecular Conformation Molecular Diagnosis Mutate Mutation Parkinsonian Disorders Patients Phase Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Play Protein Array Protein Dephosphorylation Protein Deregulation Protein Family Protein phosphatase Proteins Proteome Regulation Regulatory Element Research Resolution Role Sampling Signal Transduction Site Somatic Cell Somatic Mutation Structure Syndrome Testing Variant arm cancer type de novo mutation developmental disease disease-causing mutation early onset exome genetic regulatory protein genome sequencing human disease inhibitor insight member molecular dynamics multidisciplinary mutant nervous system disorder neurodevelopment phosphoric diester hydrolase protein phosphatase 2A regulatory subunit 65 kDa response scaffold simulation single molecule single-molecule FRET whole genome

项目摘要

PROJECT SUMMARY Protein phosphatase 2A (PP2A) is a major Ser/Thr phosphatase with complex regulation and composition. Deregulation of PP2A holoenzymes leads to devastating human diseases, including multiple types of cancer and neurological disorders. In recent years, whole exome/genome sequencing for the molecular diagnosis identified broad disease mutations in PP2A subunits and substrates in cancer and neurological disorders. The diagnosis of de novo PP2A mutations in developmental disorders enthused unprecedented multidisciplinary phosphatase research and interactions with patient families. De novo mutations in several members of B56 family of PP2A regulatory subunits, such as B56δ and B56γ, cause neurological disorders, known as Jordan Syndrome. The same somatic mutations were found in cancer patients. Built on our preliminary cryo-EM, molecular dynamic simulations, biochemical, single molecule, reverse phase protein array, and cell biology studies, here we aim to decipher the mechanisms of B56 disease mutations in cancer and intellectual disabilities. We will determine the cryo-EM structures of WT and disease variants of the PP2A-B56δ holoenzyme (Specific Aim 1), investigate how intellectual disability (ID) mutations alter holoenzyme dynamics, activation phosphorylation, and interactions with regulatory proteins, combining biochemical dissections, advanced molecular dynamic simulation approaches, and single-molecule fluorescence resonance energy transfer (smFRET) (Specific Aim 2), and test our hypothesis on the discriminating and merging mechanisms of B56δ and B56γ ID mutations in perturbing the cAMP negative feedback signaling via MAPK and CREB signaling, respectively (Specific Aim 3). These studies will shed light on a highly dynamic regulation machinery involving the folding of long disordered regions against the holoenzyme core and a super-long dynamic interface harboring the majority of residues mutated in Jordan Syndrome, multiple activation phophorylation sites, and regulatory elements that suppress both the phosphatase active site and the substrate-binding groove.

项目摘要蛋白质磷酸酶2a（PP2A）是具有复杂调节和组成的主要Ser/THR磷酸酶。 PP2A全酶的放松管制会导致毁灭性的人类疾病，包括多种类型的癌症和神经系统疾病。近年来，确定的分子诊断的整个外显/基因组测序癌症和神经系统疾病的PP2A亚基和底物中的广泛疾病突变。诊断发育障碍中的从头pp2a突变，富有前所未有的多学科磷酸酶研究和与患者家庭的互动。 B56 pp2a家族的几个成员的从头突变调节亚基，例如B56δ和B56γ，引起神经系统疾病，称为约旦综合征。在癌症患者中发现了相同的体细胞突变。基于我们的初步冷冻EM分子动态模拟，生化，单分子，反相蛋白阵列和细胞生物学研究，我们的目的是破译癌症和智力残疾中B56疾病突变的机制。我们将确定 pp2a-b56Δ全酶的WT和疾病变异的冷冻EM结构（特定目标1），研究如何如何研究智力残疾（ID）突变改变了全酶动力学，激活磷酸化和与调节性蛋白质，结合生化解剖，晚期分子动态模拟方法，和单分子荧光共振能量转移（SMFRET）（特定AIM 2），并测试我们的假设关于B56δ和B56γID突变的区分和合并机制在扰动CAMP负面分别通过MAPK和CREB信号传导反馈信号（特定目标3）。这些研究将浮出水面在高度动态的调节机械上，涉及长期无序区域的折叠 Holoenzyme Core和一个超长的动态界面，该界面具有约旦突变的大多数残留物综合征，多种激活哲学位点和调节元素，这些元素抑制了这两种磷酸酶活性位点和底物结合凹槽。