STRUCTURAL BIOLOGY OF MEMBRANE PROTEINS

膜蛋白的结构生物学

• 批准号: 7598561
• 负责人: MICHAEL G MALKOWSKI
• 金额: $ 1.62万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598561
• 关键词: Animals; Carbon; Caribbean natives; Caribbean region; Cells; Class; Computer Retrieval of Information on Scientific Projects Database; Condition; Cytochromes; Data Collection; Data Set; Detergents; Environment; Enzymes; Freezing; Funding; Grant; Human; Institution; Invertebrates; Laboratories; Lipoxygenase; Membrane; Membrane Proteins; Mus; Oryza sativa; Oxygen; Oxygenases; Plants; Polyunsaturated Fatty Acids; Production; Prostaglandin-Endoperoxide Synthase; Research; Research Personnel; Resolution; Resources; Roentgen Rays; Source; United States National Institutes of Health; Work; coral; cyclooxygenase 1; cyclooxygenase 2; lipid mediator; pathogen; research study; structural biology; synchrotron radiation

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. The research focus in the Malkowski laboratory is to understand how different classes of integral membrane enzymes utilize stereoselective oxygenations to generate unique oxylipins from a defined set of polyunsaturated fatty acid (PUFA) substrates. Oxylipins are bioactive lipid mediators found in plants and animals that are biosynthesized from 18-22 carbon PUFAs through the addition of one, two, or four molecules of oxygen via the catalytic activity of cytochrome P450s, lipoxygenases (LOX), and cyclooxygenases (COX), respectively. Diffraction studies are focused on three different integral membrane enzymes involved in oxylipin production in plants, animals, and invertebrates: a) pathogen-inducible oxygenase (PIOX) from Oryza sativa; b) human and murine cyclooxygenase-2 (COX-2); and c) a cyclooxygenase enzyme from the Caribbean coral Plexaura homomalla. This project requires synchrotron radiation for two major reasons. First, while crystals of COX-2 grow large, they diffract only weakly to 8-10 A¿` on laboratory X-ray sources. Second, their relatively empty unit cell, combined with their sensitivity to any change in the detergent environment, make handling these crystals for data collection difficult. Often many crystals must be screened to find one that maintains minimal mosaicity (<2 ¿`) and reasonable diffraction. Experiments carried out at CHESS on the A1 line in November 2006 allowed us to work out freezing conditions that were amenable to obtaining full datasets of COX-2, with max resolutions between 4-2.7A.

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