Protein interactions by analytical ultracentrifugation

通过超速离心分析蛋白质相互作用

• 批准号: 7210453
• 负责人: PETER G.W. GETTINS
• 金额: $ 33.42万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-22 至 2008-03-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7210453
• 关键词: Address; Angiogenesis Inhibitors; Antioxidants; Antithrombins; Bacillus anthracis; Biochemistry; Biotechnology; Blood coagulation; Caspase; Chicago; Class; Complement; Complex; Dentistry; Detection; Development; Disease; Dissociation; Endopeptidases; Enzymes; Equipment; Factor Xa; Fluorescence; Funding; HIV; HIV Envelope Protein gp120; Housing; Human; Illinois; Infection; Knowledge; LDL-Receptor Related Proteins; Ligands; Measurable; Measures; Medicine; Metabolic Pathway; Molecular Biology; Molecular Chaperones; Mouth Diseases; Multiple Partners; Pan Genus; Peptide Hydrolases; Pharmacologic Substance; Pharmacy facility; Physiological Processes; Process; Prodrugs; Protease Inhibitor; Proteins; Range; Regulation; Research; Role; SARS Coronavirus Protease Pathway; SARS coronavirus; Sampling; Scaffolding Protein; Serpins; Severe Acute Respiratory Syndrome; Site; Solutions; System; Therapeutic; Universities; Viral Envelope Proteins; Virus Diseases; alpha 2-Glucoproteins; analytical ultracentrifugation; college; design; enthalpy; instrumentation; interest; novel; plasma protein Z; protein protein interaction; protein structure function; response; structural biology

DESCRIPTION (provided by applicant): The Center for Structural Biology at the University of Illinois at Chicago (UIC) proposes acquisition of a Beckman Coulter XL-I analytical ultracentrifuge, with accessories, including the newly-available Fluorescence Detection System from Aviv Biomedical, to advance the research of five NIH-funded research groups, all of which involve the study of protein interactions with other proteins or ligands.The equipment will be housed in the Center for Structural Biology in a shared equipment room. The primary users belong to three departments (Biochemistry, Center for Pharmaceutical Biotechnology, and the Center for Molecular Biology of Oral Disease) in three colleges (Medicine, Pharmacy and Dentistry) of the university. The equipment will complement currently available shared instrumentation that includes ultra high field NMR, fluorescence, and CD spectrometers and DSC and ITC calorimeters by enabling quantitative examination of the solution interactions of proteins of interest with other proteins and ligands, irrespective of whether there are spectroscopic changes or measurable enthalpy changes in the interactions. Fluorescence detection capabilities will significantly extent the capabilities of the XL-I in terms of the range of systems that can be examined, including range of dissociation constants that can be measured, the complexity of systems analyzable, and sample sparing. Systems that will be studied include (i) viral envelope proteins gp41 and gp120 from HIV and the S1 and S2 from SARS coronavirus (Caffrey), (ii) interactions of the LDL receptor related protein with its chaperone RAP and with serpin-proteinase ligands, and characterization of domain-domain interactions in the human pan-proteinase inhibitor alpha-2-macroglobulin (Gettins), (iii) protein-protein interactions in the regulation of pro-drug processing enzymes, and in the role of scaffold proteins in organizing multiple partners at specific cellular sites (Lavie), (iv) heterogeneous interactions in the antioxidant response, the oligomerization state of class 3 deacetylases, of Bacillus anthracis metabolic pathway enzymes, and of proteases from the SARS coronavirus that will be targeted for inhibition by development of novel protease inhibitors (Mesecar) and (v) conformational changes in antithrombin in the antiangiogenic forms, organization of crmA-caspase complexes, and studies on the complex formed between ZPI, protein Z and factor Xa that permits inhibition of factor Xa by ZPI (Olson). The research to be carried out using this equipment addresses basic questions of the structure and function of proteins that are involved in human viral infections such as HIV or SARS, or normal physiological processes such as blood coagulation. The knowledge gained will be invaluable for design of therapeutics against such infections or in amelioration of diseases associated with aberrant functioning of processes such as blood coagulation.

描述（申请人提供）：位于芝加哥的伊利诺伊大学结构生物学中心（UIC）提议收购Beckman Coulter XL-I分析超级离心机及其配件，包括Aviv Biomedical新推出的荧光检测系统，以推进NIH资助的五个研究小组的研究，所有这些都涉及蛋白质与其他蛋白质或配体相互作用的研究。2这些设备将被安置在结构生物学中心的一个共用设备室里。主要用户属于该大学三个学院（医学、药学和牙科）的三个系（生物化学、药物生物技术中心和口腔疾病分子生物学中心）。该设备将补充目前可用的共享仪器，包括超高场NMR，荧光和CD光谱仪以及DSC和ITC量热仪，通过定量检查感兴趣的蛋白质与其他蛋白质和配体的溶液相互作用，无论相互作用中是否存在光谱变化或可测量的焓变化。荧光检测能力将大大扩展XL-I在可检测系统范围方面的能力，包括可测量的解离常数范围、可分析系统的复杂性和样品保留。将研究的系统包括（i）来自HIV的病毒包膜蛋白gp 41和gp 120以及来自SARS冠状病毒的S1和S2（Caffrey），（ii）LDL受体相关蛋白与其伴侣RAP和丝氨酸蛋白酶抑制剂蛋白酶配体的相互作用，以及人泛蛋白酶抑制剂α-2-巨球蛋白（Gettins）中结构域-结构域相互作用的表征，（iii）前药加工酶的调节中的蛋白质-蛋白质相互作用，以及支架蛋白在特定细胞位点组织多个配偶体中的作用（Lavie），（iv）抗氧化反应中的异质相互作用，炭疽芽孢杆菌代谢途径酶的3类脱乙酰酶的寡聚化状态，和SARS冠状病毒的蛋白酶，其将通过开发新的蛋白酶抑制剂（Mesecar）而被靶向抑制，和（v）抗血管生成形式的抗凝血酶的构象变化，crmA-caspase复合物的组织，以及对ZPI、蛋白Z和因子Xa之间形成的复合物的研究，该复合物允许ZPI抑制因子Xa（Olson）。使用该设备进行的研究解决了与人类病毒感染（如HIV或SARS）或正常生理过程（如血液凝固）有关的蛋白质结构和功能的基本问题。所获得的知识对于设计针对此类感染的治疗方法或改善与诸如血液凝固的过程的异常功能相关的疾病将是非常宝贵的。