HUMAN MESOTRYPSIN S195A - COMPLEX WITH APPI - CRYSTAL SCREEN CONDITION #33

人间胰蛋白酶 S195A - 与 APPI 复合物 - 晶体筛选条件

• 批准号: 8170674
• 负责人: Evette S Radisky
• 金额: $ 0.24万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170674
• 关键词: Active Sites; Affinity; Alzheimer' s Disease; Amino Acids; Aprotinin; Arginine; Binding; Brain; Cleaved cell; Complex; Computer Retrieval of Information on Scientific Projects Database; Development; Docking; Drug Delivery Systems; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzyme Precursors; Enzymes; Funding; Glycine; Grant; Human; Institution; Isoenzymes; Laboratories; Malignant Neoplasms; Minor; Multiple Sclerosis; Myelin Basic Proteins; Pancreas; Pancreatitis; Play; Process; Protein Precursors; Proteins; Relative (related person); Research; Research Personnel; Resistance; Resources; Roentgen Rays; Role; SPINK1 gene; Source; Soybeans; Specificity; Structure; Study models; Tissues; Trypsin; Trypsin Inhibitors; United States National Institutes of Health; Work; base; drug development; human disease; inhibitor/antagonist; insight; mesotrypsin; preference

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. There are three human trypsin isozymes: cationic trypsin, anionic trypsin, and mesotrypsin. All three are produced and secreted by the pancreas as digestive proenzymes, and are also produced to a lesser extent in other tissues. Trypsins are typically regulated through strong inhibition by endogenous small protein inhibitors, but the minor isozyme mesotrypsin is uniquely resistant to inhibition, binding with reduced affinity to small protein inhibitors and degrading them as substrates. The inhibitor-degrading activity of mesotrypsin has been postulated to play a role in the development of pancreatitis, through clearance of the protective pancreatic trypsin inhibitor SPINK1. Mesotrypsin is also highly expressed in the brain, and has been postulated to play a role in the processing of myelin basic protein in the pathological mechanism of multiple sclerosis. In addition, preliminary work in our own laboratory suggests that mesotrypsin plays a role in progression to malignancy in several cancers. A single amino acid change in the enzyme active site, substitution of arginine for a conserved glycine, appears to be wholly responsible for the resistance of mesotrypsin to protein inhibitors. Modeling studies based on existing X-ray structures for several trypsin/inhibitor complexes and the structure of uncomplexed mesotrypsin indicate that this arginine residue interferes with inhibitor docking in the active site, and that substantial conformational changes must occur at the enzyme/inhibitor interface before mesotrypsin can bind to a protein inhibitor. The relative binding affinities of mesotrypsin to a variety of trypsin inhibitors do not parallel the preferences of other trypsins, indicating that mesotrypsin/inhibitor binding energetics may be dominated by different interactions than those that determine the specificities of more well-studied trypsins. It is our aim to solve the structures of human mesotrypsin and human cationic trypsin in complex with several protein inhibitors, including bovine pancreatic trypsin inhibitor (BPTI), Alzheimer precursor protein inhibitor domain (APPI), and soybean trypsin inhibitor (SBTI). We expect that comparison of these structures (1) will reveal the conformational changes that allow mesotrypsin to bind inhibitors, (2) will give clues to explain why mesotrypsin, in contrast to cationic trypsin, is then able to cleave bound inhibitors, and (3) will give insight into the different specificities of mesotrypsin and cationic trypsin. The involvement of mesotrypsin in a number of human diseases makes it a potential drug target, and a structural understanding of the inhibitor resistance of this enzyme will facilitate efforts towards drug development.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 有三个人胰蛋白酶同工酶：阳离子胰蛋白酶，阴离子胰蛋白酶和中戊丙蛋白酶。 这三个是由胰腺作为消化性酶产生和分泌的，并且在其他组织中也较小生产。 胰蛋白酶通常通过内源性小蛋白抑制剂的强抑制来调节，但是次要同工生丙吡啶蛋白酶对抑制具有独特的耐药性，与对小蛋白质抑制剂的亲和力降低并将其降解为底物。 通过清除保护性胰腺胰蛋白酶抑制剂Spink1，据推测，间替蛋白的抑制剂降解活性在胰腺炎的发展中发挥作用。 美努丁蛋白酶在大脑中也高度表达，并被认为在多发性硬化症的病理机理中髓磷脂碱性蛋白质的加工中发挥作用。 此外，在我们自己的实验室中的初步工作表明，中丙蛋白酶在几种癌症的恶性肿瘤发展中起作用。 酶活性位点中的单个氨基酸变化，将精氨酸取代为保守的甘氨酸，似乎完全负责间替型蛋白酶对蛋白质抑制剂的抗性。 基于几种胰蛋白酶/抑制剂复合物的现有X射线结构的建模研究以及未复杂的间替蛋白蛋白的结构表明，这种精氨酸残基会干扰活性部位中的抑制剂对接，并且必须在酶促蛋白与蛋白质中结合的酶/抑制剂界面处发生实质性构型变化。 间替肾蛋白与多种胰蛋白酶抑制剂的相对结合亲和力与其他胰蛋白酶的偏好不相似，这表明间替型胰蛋白酶/抑制剂结合能的相互作用可能以不同的相互作用为主，而不是确定更精心培训的胰蛋白酶的特异性。 我们的目的是与几种蛋白抑制剂（包括牛胰腺胰蛋白酶抑制剂（BPTI），alzheimer前体蛋白抑制剂结构域（APPI）和大豆胰蛋白酶抑制蛋白蛋白酶抑制剂（SBTI）（SBTI），我们的目的是与几种蛋白质抑制剂（包括牛胰腺胰蛋白酶抑制剂（BPTI），Alzheimer前体蛋白抑制剂结构域（BPTI）和牛皮蛋白酶抑制蛋白抑制剂（BPTI）和牛皮蛋白酶抑制剂（SBTI），解决了人类性丙烯蛋白酶和人阳离子胰蛋白酶的结构。 我们预计这些结构（1）的比较将揭示构象变化，构象变化允许中共磷酸蛋白结合抑制剂，（2）将提供线索来解释为什么与阳离子胰蛋白酶相反，与阳离子胰蛋白酶相反，然后能够切断绑定的抑制剂，并且（3）将洞悉耶和华蛋白酶的不同特异性。 间替统蛋白在许多人类疾病中的参与使其成为潜在的药物靶标，并且对该酶抑制剂抗性的结构理解将有助于为药物开发的努力。