SINGLE CRYSTAL X-RAY DATA COLLECTION FOR PHOSPHOLIPASE AND INHIBITORS

磷脂酶和抑制剂的单晶 X 射线数据收集

• 批准号: 7601592
• 负责人: WILLIAM H CHURCH
• 金额: $ 0.55万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601592
• 关键词: Ascaridil; Cell physiology; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytosolic Phospholipase A2; Data; Data Collection; Data Set; Dinoprostone; Disease; Enzymes; Fibroblasts; Funding; Grant; HTATIP gene; Housing; Inflammatory; Institution; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Numbers; PLA2G4A gene; Pathway interactions; Phospholipase A2; Production; Prostaglandin-Endoperoxide Synthase; Proteins; Research; Research Personnel; Resolution; Resources; Source; Structure; Synovial Cell; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; United States National Institutes of Health; Up-Regulation; cancer cell; cyclooxygenase 1; cyclooxygenase 2; cytokine; density; enzyme activity; human PLA2G4A protein; human TNF protein; inhibitor/antagonist; insight; phospholipase inhibitor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Secreted phospholipase A2 (sPLA2) enzymes regulate cytokine-mediated inflammatory pathways. Exogenous Group IIA sPLA2 (sPLA2-IIA) can enhance tumour necrosis factor (TNF)-stimulated prostaglandin E2 (PGE2) production via the upregulation of NF-B-dependent cyclooxygenase(COX) protein levels in fibroblast-like synovial cells from RA patients1. The activity of cytoplasmic PLA2-(cPLA2-) is required. Proliferation of prostate cancer cells induced by exogenous sPLA2-IIA requires both sPLA2-IIA enzyme activity and cPLA2- activity2. Thus, exogenous sPLA2-IIA modulates cell function by distinct context-dependent mechanisms. We have developed inhibitors that block both the enzymatic and activity independent actions of sPLA2-IIA3 requiring the elucidation of the structure of sPLA2-IIA complexed with our inhibitors. We have grown several different crystal forms of sPLA2-IIA4 in the presence of our inhibitors, a number of which have been solved and refined from several 2.8 Ǻ datasets collected in-house. There is strong evidence of our inhibitors complexed with sPLA2-IIA, however the inhibitor density is not sufficiently well-resolved to determine the precise interactions of our inhibitors from current data. We require high precision, high resolution datasets in order to better define the structure and interactions of the inhibitors and in turn provide valuable insight into the mechanism by which sPLA2-IIA exerts its effects in inflammatory diseases and prostate cancer. 1Bidgood, M.J. et al(2000) J. Immunol. 165, 2790;2Sved, P. et al(2004) Cancer Res. 64, 6934; 3Church, W.B. et al(2001), J. Biol. Chem. 276, 33156; 4Church, W.B. et al(2000),Acta Cryst. D56,1482

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 分泌型磷脂酶A2（sPLA 2）酶调节精氨酸介导的炎症途径。外源性IIA族sPLA 2（sPLA 2-IIA）可通过上调来自RA患者的成纤维细胞样滑膜细胞中NF-β依赖性环氧合酶（考克斯）蛋白水平来增强肿瘤坏死因子（TNF）刺激的前列腺素E2（PGE 2）产生1。 需要胞质PLA 2-（cPLA 2-）的活性。由外源性sPLA 2-IIA诱导的前列腺癌细胞增殖需要sPLA 2-IIA酶活性和cPLA 2-活性2。因此，外源性sPLA 2-IIA通过不同的环境依赖性机制调节细胞功能。我们已经开发了阻断sPLA 2-IIA 3的酶促和活性独立作用的抑制剂，需要阐明与我们的抑制剂复合的sPLA 2-IIA的结构。我们已经在我们的抑制剂的存在下生长了sPLA 2-IIA 4的几种不同的晶体形式，其中一些已经从内部收集的几个2.8数据集中解决和改进。有强有力的证据表明我们的抑制剂与sPLA 2-IIA复合，然而，抑制剂密度不足以很好地分辨，以根据当前数据确定我们的抑制剂的精确相互作用。我们需要高精度，高分辨率的数据集，以更好地定义抑制剂的结构和相互作用，并反过来提供有价值的洞察sPLA 2-IIA在炎症性疾病和前列腺癌中发挥作用的机制。 1Bidgood，M.J.等（2000）J.Immunol.165，2790;2Sved，P.等（2004）Cancer Res.64，6934; 3Church，W.B.等人（2001），J. Biol. Chem. 276，33156; 4Church，W.B.等人（2000），Acta Cryst. D56，1482