PH domains as Calmodulin binding domains

PH 结构域作为钙调蛋白结合结构域

• 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
• 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

 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（由适用提供）：磷酸肌醇3-激酶（PI 3-激酶，PI3K）途径传达对淋巴细胞功能至关重要的信号。 PI3K在质膜上生成磷脂PIP3，以募集包含Pleckstrin同源（pH）结构域的蛋白质。 pH结构域是一种在从酵母到哺乳动物的生物体中表达的蛋白质中发现的进化结构褶皱。 pH结构域的核心是一个7链β桶，由大约120个氨基酸编码，由两个抗平行β片和一个C末端α螺旋组成。哺乳动物基因组编码在蛋白质中发现的大约300个pH结构域，这些蛋白质发挥了分类功能，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结构域与具有高特异性和亲和力的磷酸肌醇（KD：NM -μm范围）结合。 pH结构域通常通过带正电的赖氨酸和精氨酸保留在基本基序KXN（K/R）XR中的磷酸肌醇相互作用。但是，并非所有pH结构域都与PIP3结合。几个pH结构域与磷酸肌醇相互作用，这些磷酸肌醇有选择地富集在其他膜室中，例如在静止的质膜上的Golfi膜中的PI4P或PIP2。这是将脂质特异性传达到pH结构域的构成的关键机制，用于在细胞内空间隔离不同的效应蛋白。最近，我们意外地将Ca2+传感蛋白钙调蛋白确定为TEC激酶，ITK和SER/THR激酶AKT的PIP3结合pH结构域的蛋白质配体。 CAM和PIP3合作以增强彼此与ITK pH结构域的结合，在T细胞中需要最佳产生促炎性细胞因子IL-17A。两个单独的pH结构域与CAM相互作用的能力使我们研究了pH结构域通常与CAM相互作用的潜力。在此R03应用程序中，我们建议系统地评估pH结构域作为CAM绑定域，以生成在线数据库，该数据库将pH域分类为脂质，CAM和/或小GTPase粘合剂。这里生成的试剂也将沉积在DNA矢量存储库中以进行一般分布。