Protein-protein interaction domains: targets for neuropharmacologic intervention

蛋白质-蛋白质相互作用域：神经药理学干预的目标

• 批准号: 7229861
• 负责人: Kevin K. Caldwell
• 金额: $ 16.21万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7229861
• 关键词: ATF2 gene; Amino Acid Sequence; Applications Grants; Arginine; Attention; Binding; Biochemistry; Biological Assay; Brain; Cancer Biology; Cell membrane; Cell physiology; Cells; Cellular biology; Charge; Complex; Consensus Sequence; Coupled; Data; Development; Differentiation and Growth; Diffusion; Docking; Fluorescein; Fluorescein-5-isothiocyanate; Fluoresceins; Future; Goals; Immunoblotting; In Vitro; Incubated; Intervention; Isoenzymes; Isothiocyanates; JNK-activating protein kinase; JUN gene; Label; Learning; MAPK10 gene; MAPK14 gene; Measures; Mediating; Memory; Mental Health; Mitogen Activated Protein Kinase 1; Mitogen-Activated Protein Kinases; Modification; Molecular; Neurobiology; Neuroglia; Neurons; Palmitates; Pathway interactions; Peptides; Phosphatidylinositols; Phospholipase C; Phosphorylation; Physiological; Play; Preparation; Process; Protein Binding; Protein Binding Domain; Protein Kinase Interaction; Proteins; Recombinants; Research; Research Personnel; Role; Signal Transduction; Site; Stimulus; Structure; Substrate Interaction; Tertiary Protein Structure; Testing; Vesicle; base; brain cell; design; extracellular; in vivo; inhibitor/antagonist; interest; programs; protein aminoacid sequence; protein protein interaction; response; seal; stress activated protein kinase; tat Protein; tool

DESCRIPTION (provided by applicant): Cells possess a variety of means to integrate the external stimuli to which they are exposed into coordinated cellular responses. Both direct and indirect interactions between the components of signal transduction systems play important roles in these processes. The discovery of sequence motifs that mediate protein- protein interactions, coupled with the availability of protein amino acid sequence data, allows for the identification of putative protein binding pairs. The studies in this grant application arose from our recognition of amino acid sequences within the primary structures of phosphatidylinositol-specific phospholipase C-y (PLC-y) and PLC-p isozymes that conform to consensus sequences of docking motifs/so-called D-domains, for mitogen-activated protein kinases (MAPKs). Our initial studies support the hypothesis that PLC-y and PLC-p isozymes directly interact with MAPKs. In the proposed studies we aim to substantiate the identification of the putative D-domains and to develop cell-permeable pharmacologic tools that can be used to study the role of PLC-MAPK protein-protein interactions in vivo. The long-term goal for this research is to understand the role of PLC-MAPK signal integration in mental health. The proposed studies consist of three aims. Aim 1 is to determine whether the identified D-domain sequences bind MAPKs in vitro. Aim 2 studies will assess the ability of the identified D-domain peptide sequences to block in vitro interactions between MAPKs and their substrates; in addition, we will determine whether modifications that will be used to promote transmembrane delivery of the D-domain peptides in Aim 3 alter their ability to disrupt MAPK-substrate interactions. In Aim 3 we will quantify the delivery of the peptides into synaptoneurosomes, a preparation of sealed vesicles derived from brain. The results obtained from the proposed studies will expand our understanding of mechanisms of signal integration, providing important information about the means through which complex extracellular stimuli are integrated into coordinated cellular responses. In addition, these studies will provide valuable pharmacologic tools that can be used for studying the role of PLC-MAPK interactions in various cellular responses, including plasticity, growth, and differentiation. This, in turn, will aid in our understanding of both normal and aberrant cell functioning. Thus, these studies will have significant impact on several research fields, including neurobiology, cell biology, biochemistry, and cancer biology.

描述（由申请人提供）：细胞拥有多种方式将其所暴露的外部刺激整合到协调的细胞反应中。信号转导系统组件之间的直接和间接相互作用在这些过程中都发挥着重要作用。介导蛋白质-蛋白质相互作用的序列基序的发现，加上蛋白质氨基酸序列数据的可用性，使得能够鉴定推定的蛋白质结合对。这项资助申请中的研究源于我们对磷脂酰肌醇特异性磷脂酶 C-y (PLC-y) 和 PLC-p 同工酶一级结构内氨基酸序列的认识，这些序列符合对接基序/所谓的 D 结构域的共有序列，用于丝裂原激活蛋白激酶（MAPK）。我们的初步研究支持 PLC-y 和 PLC-p 同工酶直接与 MAPK 相互作用的假设。在拟议的研究中，我们的目标是证实假定的 D 结构域的鉴定，并开发可用于研究体内 PLC-MAPK 蛋白质-蛋白质相互作用的作用的细胞渗透性药理学工具。这项研究的长期目标是了解 PLC-MAPK 信号整合在心理健康中的作用。拟议的研究包括三个目标。目标 1 是确定所鉴定的 D 结构域序列是否在体外结合 MAPK。目标 2 研究将评估已识别的 D 结构域肽序列阻断 MAPK 与其底物之间体外相互作用的能力；此外，我们将确定用于促进 Aim 3 中 D 结构域肽跨膜递送的修饰是否会改变其破坏 MAPK-底物相互作用的能力。在目标 3 中，我们将量化肽进入突触神经体（一种源自大脑的密封囊泡的制剂）的递送。从拟议的研究中获得的结果将扩大我们对信号整合机制的理解，提供有关复杂的细胞外刺激整合到协调的细胞反应中的方法的重要信息。此外，这些研究将提供有价值的药理学工具，可用于研究 PLC-MAPK 相互作用在各种细胞反应中的作用，包括可塑性、生长和分化。反过来，这将有助于我们了解正常和异常的细胞功能。因此，这些研究将对神经生物学、细胞生物学、生物化学和癌症生物学等多个研究领域产生重大影响。

项目成果

Identification of mitogen-activated protein kinase docking sites in enzymes that metabolize phosphatidylinositols and inositol phosphates.

鉴定代谢磷脂酰肌醇和磷酸肌醇的酶中丝裂原激活的蛋白激酶对接位点。

• 发表时间: 2006-01-30
• 作者: Caldwell, Kevin K;Sosa, Marcos;Buckley, Colin T
• 通讯作者: Buckley, Colin T