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PH domains as Calmodulin binding domains

作为钙调蛋白结合域的 PH 域

基本信息

批准号：
9199208
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/9199208
关键词：
1-Phosphatidylinositol 3-Kinase Address Affinity Amino Acids Arginine Binding Binding Proteins Binding Sites C-terminal Calcium ion Calmodulin Calmodulin-Binding Proteins Cell Cycle Regulation Cell Survival Cell membrane Cells Charge Communities Complement Cytoskeletal Modeling DNA Data Databases Deposition Family Golgi Apparatus Immune response In Vitro Inflammatory Interferometry Interleukin-17 Length Ligand Binding Ligands Lipid Binding Lipids Liposomes Luciferases Lymphocyte Function Lysine 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 T-Lymphocyte TEC Protein Tyrosine Kinase Tertiary Protein Structure Therapeutic Validation Yeasts alpha helix base beta pleated sheet cytokine genetic regulatory protein immunological synapse 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。 PI3K 在质膜上生成磷脂 PIP3，以招募含有 pleckstrin 同源 (PH) 结构域的蛋白质。 PH 结构域是一种进化上保守的结构折叠，存在于从酵母到哺乳动物等生物体中表达的蛋白质中。 PH结构域的核心是7链β-桶，由大约120个氨基酸编码，由两个反向平行的β折叠和C端α螺旋组成。哺乳动物基因组编码大约 300 个蛋白质中的 PH 结构域，这些蛋白质执行多种功能，包括细胞激活、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磷酸肌醇相互作用。然而，并非所有 PH 域都与 PIP3 结合。多个 PH 结构域与选择性富集在其他膜区室中的磷酸肌醇相互作用，例如高尔基膜内的 PI4P 或静息质膜上的 PIP2。因此，将脂质特异性传递到 PH 结构域构成了细胞内空间隔离不同效应蛋白的关键机制。最近，我们出乎意料地发现 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 所必需的。两个独立的 PH 结构域与 CaM 相互作用的能力促使我们研究 PH 结构域与 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 结构域作为 CaM 结合结构域，以便生成将 PH 结构域分类为脂质、CaM 和/或小 GTP 酶结合物的在线数据库。这里生成的试剂也将被存入 DNA 载体存储库中以供普遍分发。