PH domains as Calmodulin binding domains

PH 结构域作为钙调蛋白结合结构域

基本信息

批准号：
9023737
负责人：
Yina Hsing Huang
金额：
$ 8.1万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9023737
关键词：
1-Phosphatidylinositol 3-Kinase Address Affinity Amino Acids Binding Binding Proteins Binding Sites C-terminal Calcium ion Calmodulin Calmodulin-Binding Proteins Cell Cycle Regulation Cell Survival Cell membrane Cells Charge Communities Cytoskeletal Modeling DNA Data Databases Deposition Family Golgi Apparatus Immune Immune response In Vitro Inflammatory Interferometry Interleukin-17 Length Ligand Binding Ligands Lipid Binding Lipids Liposomes Luciferases Lymphocyte Function Mammals Measures Mediating Membrane Monomeric GTP-Binding Proteins Mus Nature Organism PH Domain Pathway interactions Peptides Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Phospholipids Phosphotransferases Production Proteins Reagent Recruitment Activity Regulation Research Resources Rest SH3 Domains Scientist Sequence Analysis Signal Pathway Signal Transduction Specificity Synapses T-Lymphocyte TEC Protein Tyrosine Kinase Tertiary Protein Structure Therapeutic Validation Yeasts alpha helix arginyllysine base beta pleated sheet cytokine genetic regulatory protein in vivo mammalian genome migration prediction algorithm public health relevance repository src Homology Region 2 Domain structural biology trafficking vector

项目摘要

DESCRIPTION (provided by applicant): The phosphoinositide 3-kinase (PI 3-kinase, PI3K) pathway transduces signals critical for lymphocyte function. PI3K generates the phospholipid PIP3 at the plasma membrane to recruit proteins that contain pleckstrin homology (PH) domains. The PH domain is an evolutionarily conserved structural fold found in proteins expressed in organisms ranging from yeast to mammals. The core of the PH domain is a 7-strand β-barrel that is encoded by approximately 120 amino acids and is composed of two anti-parallel β sheets and a C-terminal α helix. The mammalian genome encodes roughly 300 PH domains found in proteins that perform diverse functions including cellular activation, cytoskeletal reorganization, vesicular trafficking, and cell cycle control. Approximately 15% of PH domains bind to phosphoinositides with high specificity and affinity (Kd: nM - low μM range). PH domains generally interact with phosphoinositides through positively charged lysine and arginine residues within the basic motif KXn(K/R)XR. However, not all PH domains bind to PIP3. Several PH domains interact with phosphoinositides that are selectively enriched in other membrane compartments, such as PI4P within the golgi membrane or PIP2 at the resting plasma membrane. Thus, conveying lipid specificity to PH domains constitutes a key mechanism for spatially sequestering distinct effector proteins within cells. Recently, we unexpectedly identified the Ca2+-sensing protein Calmodulin as a protein ligand for the PIP3-binding PH domains of the Tec kinase, Itk and the Ser/Thr kinase Akt. CaM and PIP3 cooperate to enhance each other's binding to the Itk PH domain and was required in T cells for optimal production of the pro-inflammatory cytokine IL-17A. The ability of two separate PH domains to interact with CaM led us to investigate a potential for PH domains in general to interact with CaM. In this R03 application, we propose to systematically evaluate PH domains as CaM binding domains for the purpose of generating an online database that categorizes PH domains as lipid, CaM, and/or small GTPase binders. The reagents generated here will also be deposited into a DNA vector repository for general distribution.

描述（由申请人提供）：磷酸肌醇 3 激酶（PI 3 激酶，PI3K）途径转导对淋巴细胞功能至关重要的信号，在质膜上生成磷脂 PIP3，以招募含有 pleckstrin 同源性 (PH) 结构域的蛋白质。 PH 结构域是在从酵母到哺乳动物等生物体中表达的蛋白质中发现的进化保守的结构折叠。 7 链 β-桶由大约 120 个氨基酸编码，由两个反向平行 β 折叠和 C 端 α 螺旋组成。哺乳动物基因组编码大约 300 个 PH 结构域，这些结构域存在于执行包括细胞激活在内的多种功能的蛋白质中。、细胞骨架重组、囊泡运输和细胞周期控制。大约 15% 的 PH 结构域以高特异性和亲和力与磷酸肌醇结合（Kd：nM）。 - 低 μM 范围）。PH 结构域通常通过基本基序 KXn(K/R)XR 内带正电荷的赖氨酸和精氨酸残基与 PIP3 相互作用。然而，并非所有 PH 结构域都与 PIP3 选择性结合。富含其他膜区室，例如高尔基膜内的 PI4P 或静息质膜上的 PIP2，从而将脂质特异性传递给 PH。最近，我们意外地发现 Ca2+ 感应蛋白钙调蛋白是 Tec 激酶、Itk 和 Ser/Thr 激酶 Akt 的 PIP3 结合 PH 结构域的蛋白配体。 CaM 和 PIP3 协同增强彼此与 Itk PH 结构域的结合，并且是 T 细胞最佳产生促炎细胞因子 IL-17A 所需的两种独立的能力。 PH 域与 CaM 相互作用使我们研究了 PH 域与 CaM 相互作用的可能性。在此 R03 应用中，我们建议系统地评估 PH 域作为 CaM 结合域，以便生成对 PH 域进行分类的在线数据库。作为脂质、CaM 和/或小 GTP 酶结合物，此处生成的试剂也将存入 DNA 载体存储库中以供一般分发。