Regional Biocontainment Laboratory (RBL) For Region V
John Keith Moffat, Professor
University Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 1UC6AI058606-01 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: The Ricketts (Regional Biocontainment) Laboratory would be constructed in a secure area leased from the Department of Energy at Argonne National Laboratory near Chicago, Illinois. It would serve the research, translational research, research training, service, and emergency response functions of the Midwest RCE (Regional Center of Excellence). The Midwestern RCE includes Region V scientists from Argonne National Laboratory, Battelle Memorial Institute, Illinois Institute of Technology, Mayo Clinic, Medical College of Wisconsin, Michigan State University, Northwestern University, Notre Dame University, Purdue University, University of Chicago, University of Illinois at Urbana-Champaign, University of Michigan, and University of Wisconsin. Intellectual contributions from over 100 scientists produced twenty-five inter-disciplinary, inter-institutional research programs selected by competitive peer review. The programs focus on studying CDC Category A Select Agents to develop new therapeutics, vaccines and diagnostic devices. Review mechanisms will provide access for NIAID supported non-RCE scientists, including proprietary research. An emergency response plan provides training and, when necessary complete commitment to emergency responders. The Ricketts Laboratory would have 54,100 square feet (27,500 net square feet (NSF)), with 8,900 NSF of BSL-2/BSL-3 molecular laboratory space and 13,300 NSF of BSL-2/BSL-3 animal research laboratories. It would have the capacity for simultaneous study of four or more different Select Agent Category A pathogens, by the research groups of five principal investigators and thirty-five staff scientists, with a total holding capacity for about 30,000 mice or other experimental animals
Project start date: 2003-09-30
Project end date: 2008-06-30
1UC6AI058606-01 (2003): $17374817
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to John Keith Moffat
BIOCARS A SYNCHROTRON STRUCTURAL BIOLOGY RESOURCE
John Keith Moffat, Professor
Biochemistry And Molecular Biologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 5U41RR007707-10 from National Center For Research Resources IRG: ZRG3
Abstract: We wish to design, construct and operate a single sector at the Advanced Photon Source (APS) under construction at Argonne National Laboratory. The sector will consist of an undulator and wiggler source, a bending magnet source, associated X-ray optical elements, beam transport, three hutches, a BL3 biohazards facility, and experimental apparatus suitable for five different styles of experiments in structural biology virus crystallography, small angle scattering, protein crystallography/multiple wavelength anomalous dispersion, time-resolved diffraction, and diffraction from microcrystals. The sector will be operated for the 29-member group of structural biologists denoted bioCARS (roughly 75% of the available beamtime) and for general users (roughly 25%). Technological research and development will concentrate on x-ray optics, including transmission optics and cooling schemes that will enable the optics to withstand the high heat loads imposed by the insertion device sources without significant degradation of their properties; storage phosphor and other detectors; sample handling devices such as liquid helium cryostats and thin film crystal mounts; software for efficient, user-friendly data acquisition, reduction and storage of both static and time-resolved data; and novel, experimental techniques. Collaborative and service projects will encompass X-ray diffraction from single crystals and from less-ordered membranes, fibers, and solutions. Particular emphasis will be placed on crystals with very large unit cells such as viruses, and on time-resolved studies on time scales down to an d including the X-ray pulse length at the APS, around 100ps. These studies exploit the unique properties of the APS its brilliance, intensity, and dedicated availability. Prior to the start-up of the APS in 1995-96, initial experiments and tests of x-ray equipment will be conducted at the National Synchrotron Light Source, on beam line X26C. BioCARS forms an integral part of the interdisciplinary Consortium for Advanced Radiation Sources, CARS. CARS consist of three universities (the University of Chicago, Northern Illinois University, and Southern Illinois University) and four national science groups in structural biology (BioCARS) , the earth and planetary sciences (GeoCARS), chemical sciences (ChemCARS) and soil and environmental science (Soil/EnviroCARS). CARS at present consist of 97 principal investigators, and is a Collaborative Access Team (CAT) which proposes to develop and operate two sectors at the APS, of which this request is for support of one. Substantial facilities, obtained with State and University support, are already in place or committed, including design, construction, x-ray, and metrology laboratories
Keywords: X ray, X ray crystallography, biomedical equipment resource, biomedical resource, cooperative study, structural biology, synchrotron bioimaging /biomedical imaging
Project start date: 1992-09-15
Project end date: 2002-08-14
5U41RR007707-10 (2001): $1566376
5U41RR007707-09 (2000): $1997007
5U41RR007707-07 (1998): $1200906
BIOCARS--A SYNCHROTRON STRUCTURAL BIOLOGY RESOURCE
John Keith Moffat, Professor
Biochemistry And Molecular Biologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 5U41RR007707-03 from National Center For Research Resources IRG: SSS
Abstract: We wish to design, construct and operate a single sector at the Advanced Photon Source (APS) under construction at Argonne National Laboratory. The sector will consist of an undulator and wiggler source, a bending magnet source, associated x-ray optical elements, beam transport, three hutches, a BL3 biohazards facility, and experimental apparatus suitable for five different styles of experiments in structural biology virus crystallography, small angle scattering, protein crystallography/multiple wavelength anomalous dispersion, time-resolved diffraction, and diffraction from microcrystals. The sector will be operated for the 29-member group of structural biologists denoted BioCARS (roughly 75% of the available beamtime) and for general users (roughly 25%). Technological research and development will concentrate on X-ray optics, including transmission optics and cooling schemes that will enable the optics to withstand the high heat loads imposed by the insertion device sources without significant degradation of their properties; storage phosphor and other detectors; sample handling devices such as liquid helium cryostats and thin film crystal mounts; software for efficient, user-friendly data acquisition, reduction and storage of both static and time-resolved data; and, novel experimental techniques. Collaborative and service projects will encompass X-ray diffraction from single crystals and from less-ordered membranes, fibers and solutions. Particular emphasis will be placed on crystals with very large unit cells such as viruses, and on time-resolved studies on time scales down to and including the x-ray pulse length at the APS, around 100 ps. These studies exploit the unique properties of the APS its brilliance, intensity, and dedicated availability. Prior to the start-up of the APS in 1995-96, initial experiments and tests of X-ray equipment will be conducted at the National Synchrotron Light Source, on beam line X26C. BioCARS forms an integral part of the interdisciplinary Consortium for Advanced Radiation Sources, CARS. CARS consist of three universities (The University of Chicago, Northern Illinois University, and Southern Illinois University) and four national scientific groups, in structural biology (BioCARS), the earth and planetary sciences (GeoCARS), chemical sciences (ChemCARS) and soil and environmental science (Soil/EnviroCARS). CARS at present consists of 97 principal investigators, and is a Collaborative Access Team (CAT) which proposes to develop and operate two sectors at the APS, of which this request is for support of one. Substantial facilities, obtained with State and University support, are already in place or committed, including design, construction, X-ray, and metrology laboratories
Keywords: X ray, X ray crystallography, biomedical equipment resource, cooperative study, particle accelerator, structural biology
Project start date: 1992-09-15
Project end date: 1997-09-14
5U41RR007707-03 (1994): $2306776
A MACROMOLECULAR DIFFRACTION BIOTECHNOLOGY RESOURCE: MAC
John Keith Moffat, Professor
Cornell University Ithaca Office Of Sponsored Programs Ithaca, Ny 148502820
Grant 5P41RR001646-05 from National Center For Research Resources IRG: SSS
Abstract: We wish to design, construct and evaluate a complete x-ray biotechnology facility at the Cornel High Energy Synchrotron Source, CHESS. The facility will enable the rapid collection of x-ray diffraction data on single crystals of macromolecules unsuitable for laboratory study. Techniques for direct phase determination which exploit the variation in atomic scattering factors near absorption edges will be explored, as will real-time diffraction experiments on membrane or solution samples. Design and construction of the apparatus will require collaboration between biophysicists and biochemists. Crystals to be studied include animal and plant viruses, viral proteins, polypeptide hormones and enzymes which recognize specific sequences in DNA.
Keywords: BIOMEDICAL RESOURCES, INSTRUMENTATION RESOURCES, CHEMISTRY, ANALYTICAL METHODS, CRYSTALLOGRAPHY, CHEMISTRY, ANALYTICAL METHODS, X-RAY STRUCTURE ANALYSIS, MOLECULES, MACROMOLECULES
Project start date: 1983-09-01
Project end date: 1988-08-31
A MACROMOLECULAR DIFFRACTION BIOTEHNOLOGY RESOURCE: MAC
John Keith Moffat, Professor
Cornell University Ithaca Office Of Sponsored Programs Ithaca, Ny 148502820
Grant 5P41RR001646-02 from National Center For Research Resources IRG: SSS
Abstract: We wish to design, construct and evaluate a complete x-ray biotechnology facility at the Cornel High Energy Synchrotron Source, CHESS. The facility will enable the rapid collection of x-ray diffraction data on single crystals of macromolecules unsuitable for laboratory study. Techniques for direct phase determination which exploit the variation in atomic scattering factors near absorption edges will be explored, as will real-time diffraction experiments on membrane or solution samples. Design and construction of the apparatus will require collaboration between biophysicists and biochemists. Crystals to be studied include animal and plant viruses, viral proteins, polypeptide hormones and enzymes which recognize specific sequences in DNA.
Keywords: BIOMEDICAL RESOURCES, INSTRUMENTATION RESOURCES, CHEMISTRY, ANALYTICAL METHODS, CRYSTALLOGRAPHY, CHEMISTRY, ANALYTICAL METHODS, X-RAY STRUCTURE ANALYSIS, MOLECULES, MACROMOLECULES
Project start date: 1983-09-01
Project end date: 1988-08-31
BioCARS: A Synchrotron Structural Biology Resource
John Keith Moffat, Professor
Biochemistry And Molecular Biologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 5P41RR007707-15 from National Center For Research Resources IRG: ZRG1
Abstract: We will continue to operate BioCARS, a nationally-available research resource for synchrotron-based macromolecular crystallography at the Advanced Photon Source, Argonne National Laboratory. The facility contains three experimental stations, one served by either an undulator or a wiggler insertion device source and two served by a bending magnet source, all of which can be used simultaneously. All stations and the associated control areas are embedded in a BL3-level biosafety facility, to permit the safe study of, for example, pathogenic human viruses. Research by users has successfully and productively concentrated on de novo structure determination by MAD phasing; on the collection of accurate, high resolution, monochromatic data; on the study of crystals with large unit cell dimensions; and on time-resolved crystallography on time scales down to a few nanoseconds. We will continue to emphasize these areas and to support the most challenging user experiments, through our experienced technical and scientific staff. In a facility that is unique nationally and one of three internationally, we will further develop time-resolved crystallography in both core and collaborative research, largely on the insertion device station. Advances in this scientific technique have led to much broader user interest in applying it to a wide variety of systems, since it offers new insights into biological mechanism and function. Upgrades will be made to the X-ray beam lines, both to replace those older components damaged by radiation and by heating and to afford smaller focal spots that are better matched to the smaller crystals that users wish to examine. Detector and data acquisition and control software will be improved, both to support collaborative projects in structural genomics that demand high data throughput, and to enable telecrystallography, in which the user can participate effectively in all aspects of the synchrotron experiment but need not be physically present at BioCARS. Strategies for improving the diffraction quality of crystals at both room and cryogenic temperatures by annealing will be implemented. In all cases, these developments are aimed at enhancing the throughput of excellent, high-resolution diffraction data and solved scientific problems, by satisfied users
Keywords: biomedical resource, structural biology
Project start date: 1992-09-15
Project end date: 2008-07-31
5P41RR007707-15 (2006): $1508919
5P41RR007707-14 (2005): $1500226
5P41RR007707-13 (2004): $1556533
Sponsored Links Excellgen http://Excellgen.com
5P41RR007707-12 (2003): $2473691
2P41RR007707-11 (2002): $2816735
3P41RR007707-11S1 (2002): $1252800
EXPLORING LIGHT-SENSING AND SIGNALING MECHANISMS OF BACTERIOPHYTOCHROMES BY C
John Keith Moffat, Associate Professor
University Of Chicago, 5801 S Ellis Ave, Chicago, Il 60637
Abstract: 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. Light is a major environmental stimulus for both prokaryotes and eukaryotes. Phytochromes are red light photoreceptors that regulate a wide range of physiological processes such as seed germination, floral induction and phototaxis in plants, fungi and bacteria. Plant phytochromes (Phy) and bacteriophytochromes (Bph) use a linear tetrapyrrole (bilin) as a chromophore and photo-convert between red-absorbing (Pr) and far-red-absorbing (Pfr) states (Rockwell et al, 2006). However, molecular and mechanistic details of photoconversion and signal transduction mechanisms remain obscure. During the last proposal (#6457) period, we have successfully conducted experiments and collected data that allow us determine four different crystal structures of the photosensory domains from three distinct bacteriophytochromes (PaBphP from P. aeruginosa and RpBphP3/RpBphP2 from R. palustris) and several mutant structures in both Pr and Pfr states. We have solved the very first Pfr structure for phytochromes of any kind. We will continue to explore structural basis of signal transduction mechanisms in bacteriophytochromes using static and time-resolved crystallography as well as small-angle X-ray scattering techniques. Our specific aims during this proposal period are 1. to determine crystal structures of full-length bacteriophytochromes (160KD) with intact sensor and effector domains (static monochromatic crystallography); 2. to cryo-trap structural intermediates during Pr/Pfr photoconversion reaction pathways (static monochromatic crystallography); 3. to probe early structural intermediates (in ps-ms time scale) during photoconversion at ambient temperature (time-resolved Laue crystallography); 4. to explore global structural changes that transmit light signal perceived by the N-terminal sensory domains to the C-terminal effector histidine kinase domain. This requires SAXS to study samples in solution, since such global structural rearrangements are often limited in crystal lattice
Keywords: Bacteria; Bilin; C-terminal; CRISP; Cell Communication and Signaling; Cell Signaling; Computer Retrieval of Information on Scientific Projects Database; Crystallographies; Crystallography; DNA Sequence Rearrangement; Data; Eukaryota; Eukaryote; Funding; Germination; Grant; Institution; Intracellular Communication and Signaling; Investigators; Length; Light; Methods and Techniques; Methods, Other; Molecular; N-terminal; NH2-terminal; NIH; National Institutes of Health; National Institutes of Health (U.S.); Organism-Level Process; Organismal Process; P. aeruginosa; P.aeruginosa; Pathway interactions; Photoradiation; Photoreceptor Cell; Photoreceptors; Photosensitive Cell; Physiologic Processes; Physiological Processes; Phytochrome; Plant Embryos; Plants; Plants, General; Prokaryotae; Prokaryotic Cells; Pseudomonas aeruginosa; Pseudomonas pyocyanea; Radiation, X-Rays; Radiation, X-Rays, Gamma-Rays; Reaction; Rearrangement; Research; Research Personnel; Research Resources; Researchers; Resources; Roentgen Rays; Sampling Studies; Seeds; Sensory; Signal Transduction; Signal Transduction Systems; Signaling; Solutions; Source; Stimulus; Structure; Techniques; Temperature; Tetrapyrroles; Time; United States National Institutes of Health; Visual Receptor; X-Radiation; X-Rays; Xrays; Zygotes, Plant; base; biological signal transduction; chromophore; eukaryotida; experiment; experimental research; experimental study; histidine kinase; mutant; pathway; plant fungi; prokaryote; protein-histidine kinase; research study; seed; sensor; structural biology
Project start date: 2009-08-01
Project end date: 2010-07-31
Budget start date: 1-AUG-2009
Budget end date: 31-JUL-2010
5P41RR007707-17_7050 (2009): $23600
TIME-RESOLVED MACROMOLECULAR CRYSTALLOGRAPHY
John Keith Moffat, Associate Professor
University Of Chicago, 5801 S Ellis Ave, Chicago, Il 60637
Grant 5R01GM036452-25 from National Institute Of General Medical Sciences
Abstract: Our understanding of reaction mechanisms in biological systems at the molecular level is presently based largely on knowledge of static, chemically- or physically-trapped structures obtained by high resolution X-ray crystallographic and NMR techniques. However, the dynamic aspects of changes in structure are critical in all chemical and biological processes, for example catalysis, ligand binding and release, and signal transduction. To explore the mechanisms of signal transduction, we have successfully conducted time-resolved crystallographic experiments with ~100 picosecond time resolution and high crystallographic resolution on light-sensitive systems, identified the structures of short-lived intermediates and characterized the overall mechanism of light-dependent signal transduction which these intermediates populate. Recently, we have determined the static crystal structures of several novel, naturally-occurring, signaling photoreceptors which are based on a modular architecture. Those containing so-called LOV or BLUF sensor domains respond to blue light, and bacteriophytochromes containing PAS-GAF-PHY domains to red/far-red light. We will now conduct time-resolved crystallographic experiments on these proteins, and on longer constructs that contain both sensor and effector (output) domains. We address the questions How is a signal generated and transmitted? How is activity controlled by light? In each, what is the mechanism of light-dependent signal transduction? Despite the high chemical and structural diversity in natural photoreceptors, we believe that general principles of signal transduction exist. Indeed, we have prepared chimeric, artificial photoreceptors in which we have made light-sensitive a normally light-inert biological activity e.g. histidine kinase or DNA binding. We will explore these artificial photoreceptors to test and expand our general principles, afford useful tools and offer new targets for crystallization. Finally, artificial photoreceptors overcome a limitation of time-resolved crystallography many interesting systems are not light-dependent. We will pursue the question How general are these approaches? Many cancers are associated with derangement of signal transduction pathways, driven by ligand binding to chemoreceptors. The pathways may be known but the mechanisms at the molecular level are not. The natural and artificial photoreceptors whose molecular mechanisms we study have parallels to, but also key differences from, chemoreceptors the thermodynamic and structural principles of signal transduction are likely to be similar in both
Keywords: ATP[{..}]protein-tyrosine O-phosphotransferase; Absorption; Address; Arabidopsis thaliana; Architecture; Bacillus subtilis; Bacteria; Behavior; Binding; Binding (Molecular Function); Biological; Biological Function; Biological Process; Cancers; Catalysis; Cell Communication and Signaling; Cell Signaling; Chemicals; Chemoreceptors; Chicago; Collaborations; Coupling; Cress, Mouse-ear; Crystallization; Crystallographies; Crystallography; DNA Binding; DNA Binding Interaction; EPH- and ELK-Related Tyrosine Kinase; EPH-and ELK-Related Kinase; EPHA8; Electronics; Engineering / Architecture; EphA8 Protein; Ephrin Type-A Receptor 8; Ephrin Type-A Receptor 8 Precursor; Esteroproteases; Event; Exhibits; GTP Phosphohydrolases; GTPases; Goals; Guanosine Triphosphate Phosphohydrolases; Guanosinetriphosphatases; HEK3; Intracellular Communication and Signaling; Kinetic; Kinetics; Knowledge; Life; Ligand Binding; Light; Light Activity; Light Exercise; Link; Malignant Neoplasms; Malignant Tumor; Methods and Techniques; Methods, Other; Molecular; Molecular Interaction; Mouse-ear Cress; Mutagenesis, Site-Directed; O element; O2 element; Oats; Organism; Output; Oxygen; P. aeruginosa; P.aeruginosa; PTK; Pathway interactions; Peptidases; Peptide Hydrolases; Photons; Photoradiation; Photoreceptor Cell; Photoreceptors; Photosensitive Cell; Plants; Plants, General; Process; Process of absorption; Progress Reports; Proteases; Protein Tyrosine Kinase; Protein Tyrosine Kinase EEK; Protein-Serine Kinase; Protein-Serine-Threonine Kinases; Protein-Threonine Kinase; Proteinases; Proteins; Proteolytic Enzymes; Pseudomonas aeruginosa; Pseudomonas pyocyanea; Radiation, X-Rays; Radiation, X-Rays, Gamma-Rays; Reaction; Reports, Progress; Resolution; Rhodopseudomonas; Roentgen Rays; Serine Kinase; Serine-Threonine Kinases; Signal Transduction; Signal Transduction Pathway; Signal Transduction Systems; Signaling; Site-Directed Mutagenesis; Site-Specific Mutagenesis; Solutions; Structural Biologist; Structure; System; System, LOINC Axis 4; Targeted DNA Modification; Targeted Modification; Techniques; Testing; Thermodynamic; Thermodynamics; Threonine Kinase; Time; Tyrosine Kinase; Tyrosine-Protein Kinase Receptor EEK; Tyrosine-Specific Protein Kinase; Tyrosylprotein Kinase; V (voltage); Visual Receptor; X-Radiation; X-Rays; Xrays; absorption; base; biological signal transduction; biological systems; chemical kinetics; chemical reaction; chromophore; design; design and construction; designing; experiment; experimental research; experimental study; fungus; gene product; guanosinetriphosphatase; histidine kinase; hydroxyaryl protein kinase; interest; kinetics (chemistry); living system; malignancy; neoplasm/cancer; novel; pathway; protein-histidine kinase; public health relevance; research study; sensor; success; tool; tyrosyl protein kinase; voltage
Relevance: Many cancers are associated with derangement of signal transduction pathways, driven by ligand binding to chemoreceptors. The pathways may be known but the mechanisms at the molecular level are not. The natural and artificial photoreceptors whose molecular mechanisms we study have parallels to, but also key differences from, chemoreceptors: the thermodynamic and structural principles of signal transduction are likely to be similar in both
Project start date: 1990-09-01
Project end date: 2012-12-30
Budget start date: 1-JAN-2010
Budget end date: 31-DEC-2010
PFA/PA: PA-07-070
5R01GM036452-25 (2010): $602328
3R01GM036452-25S1 (2010): $105409
5R01GM036452-22 (2007): $526833
5R01GM036452-21 (2006): $548427
2R01GM036452-20 (2005): $578922
Sponsored Links Excellgen http://Excellgen.com
A MACROMOLECULAR DIFFRACTION RESOURCE: MACCHESS
John Keith Moffat, Professor
Cornell University Ithaca Office Of Sponsored Programs Ithaca, Ny 148502820
Grant 5P41RR001646-07 from National Center For Research Resources IRG: SSS
Abstract: Development of a macromolecular diffraction resource at CHESS, the Cornell High Energy Synchrotron Source will be continued. This resource devises novel x-ray diffraction techniques and apparatus, suitable for the study of macromolecules and macrmolecular assemblies such as enzymes, hormones, immunoglobulins, membranes and membrane components, DNA and DNA-protein complexes, ribosomes, and plant, insect and mammalian viruses, in the form of single crystals, fibers, membranes and solutions. Techniques exploit the unique features of a synchrotron x-ray source its intensity, its polychromatic nature, and its time structure. The Laue technique for time-resolved crystallography will be developed further using both a new focussed white beam line and a high energy undulator; novel x-ray optical components will be devised; a new form of sensitive, low noise x-ray detector, the Kodak storage phosphor detector, will be evaluated; and an x-ray wiggler beam line optimized for macromolecular crystallography, especially of viruses, will be installed. The last will be incorporated into a Biohazards Facility suitable for handling concentrated solutions and crystals of Pathogenic viruses at the BL3 level in the NIH/CDC classification. To our knowledge, it will be unique in the world. These facilities will continue to be made widely available to scientists from university, government and industry who are interested in macromolecular diffraction as a probe of structure and function.
Keywords: BIOMEDICAL RESOURCES, INSTRUMENTATION RESOURCES, CHEMISTRY, ANALYTICAL METHODS, CRYSTALLOGRAPHY, CHEMISTRY, ANALYTICAL METHODS, X-RAY STRUCTURE ANALYSIS, MOLECULES, MACROMOLECULES
Project start date: 1983-09-01
Project end date: 1993-08-31
BIOCARS: A SYNCHROTON STRUCTURAL BIOLOGY RESOURCE
John Keith Moffat
University Of Chicago, 5801 S Ellis Ave, Chicago, Il 60637
Grant 5P41RR007707-18 from National Center For Research Resources
Abstract: BioCARS is a nationally-available research resource for synchrotron-based structural biology at Sector 14 of the Advanced Photon Source, Argonne National Laboratory, that exploits the distinctive features of BioCARS and complements other structural biology beamlines at the APS and nationally. Our chief scientific emphasis lies on time-resolved crystallography on time scales from 100 ps to s, in which a substantial collaborative user community is developing. Our two experimental stations are embedded in a BSL-3 facility to allow safe study of biohazards. Both the time-resolved and biohazards capabilities are unique in the USA. The recently-upgraded insertion device beamline 14-ID is served by two dissimilar, collinear undulator sources that deliver tightly-focused, intense, tunable, monochromatic or pink X-ray beams to the 14-ID-B station, suitable for both time-resolved and standard monochromatic experiments. The bending magnet beamline 14-BM is particularly suited to study crystals with large unit cells, or at ultra-high resolution. A laser laboratory houses ps and ns pulsed lasers and other optical instrumentation essential to the time-resolved experiments. Core research and development concentrates on time-resolved crystallography. We will implement new methods for exploring irreversible reactions based on pixel array detectors, on a novel use of an existing image plate detector and on crystal rotation geometry. We will develop a new capability to study small to microcrystals with both monochromatic and Laue techniques, and examine the ultrafast time course of radiation damage at room and cryo temperatures. The small to microcrystal capability supports broadly other core R&D, collaborative and service activities. Collaborative research focuses on time-resolved crystallography and thus drives our core R&D activities. Service emphasizes study of biohazards, both as single crystals (often with large unit cells) and where feasible, as fibers or less-ordered samples; and of small to microcrystals. We will continue our high levels of user training, and of dissemination to both expert and non-expert, wider audiences, by electronic, print and face-to-face approaches. In all cases, we assist users in solving transient and static, atomic-level structures of direct relevance to significant biomedical problems. Some of these structures will be of biohazards classified at the BSL-3 level; examples include human pathogenic viruses such as West Nile virus and prions. Others such as anthrax related toxins or enzymes are key to the metabolism of pathogenic microorganisms. They are both of substantial public health interest in their own right and may yield information directed at responses to bioterrorism. Yet others, particularly the transient structures, provide a novel view of mechanisms of action, and thus underlie the development of more effective therapeutic agents
Project start date: 1997-07-01
Project end date: 2013-07-31
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
5P41RR007707-18 (2010): $1076363
5P41RR007707-17 (2009): $1205429
3P41RR007707-17S1 (2009): $342121
2P41RR007707-16A1 (2008): $1257251
TIME RESOLVED MACROMOLECULAR CRYSTALLOGRAPHY
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5R37GM036452-13 from National Institute Of General Medical Sciences IRG: BBCB
Abstract: An understanding of mechanism in biological systems at the molecular level is based both on static structure and on changes in structure. The dynamic aspects of changes in structure during processes such as enzyme catalysis, photocycling in light-sensitive systems, ligand binding and release, and protein unfolding and refolding are critical. However, the rates of interconversion of structures under physiological conditions are large, and the lifetimes of transient structures correspondingly small. The observation of short-lived, transient structures by x-ray crystallographic techniques has therefore depended on the development and application of new techniques in several areas; 1) the rapid acquisition in a time-dependent manner of accurate x-ray data generated by intense synchrotron x-ray sources; 2) the uniform, synchronous, initiation of a structural reaction in the molecules in a crystal; 3) the ability to simultaneously monitor an optical property of the crystal such as absorbance, thus yielding both an optical and an x-ray measure of progress along the reaction coordinate; and 4) the ability to process the x-ray data to yield both the time-dependent difference in structure between that at a particular instant on the reaction coordinate and that of the reactants, and (under favorable circumstances) the time-independent structures through which the system progressively evolves. in turns out that it is substantially easier to study these slow structural reactions in crystals which occur on non- physiological time scales of tens of seconds or minutes. We concentrate here on much faster structural reactions which occur on sub-second time scales, down to times as short as 100 picoseconds when the new Advanced Photon Source comes into operation at Argonne National Laboratory. These factor structural reactions occur in the time scale where heat transfer processes are important. Great care has to be taken in the experimental design to minimize the photochemical and thermal gradients which may be introduced during laser initiation of the reaction, or through irradiation with an intense, polychromatic x-ray beam. Systems to be studied include photoactive yellow protein, which undergoes a photocycle and appears to be a simple, soluble bacterial photosensor; the carbon monoxide complexes of heme proteins such as myoglobin and hemoglobin; and other proteins which are either naturally photosensitive or can be made so by employing molecules such as photoactivatable substrates. Examples of the former include rhodopsin, and of the latter, restrictocin.
Keywords: X ray crystallography, method development, molecular dynamics, photochemistry, protein structure /function, time resolved data, carboxyhemoglobin, chemical binding, chemical kinetics, conformation, heme, hemoglobin, mathematical model, myoglobin, nonvisual photoreceptor, pancreatic ribonuclease, photoactivation, photolysis, rhodopsin, structural biology, temperature, thermodynamics, laser
Project start date: 1990-09-01
Project end date: 1999-11-30
5R37GM036452-13 (1998): $295856
5R37GM036452-14 (1999): $305175
5R37GM036452-12 (1997): $285922
6TH INTL CONF ON BIOPHYSICS AND SYNCHROTRON RADIATION
John Keith Moffat, Professor
Radiologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 1R13RR013462-01 from National Center For Research Resources IRG: ZRR1
Abstract: Adapted from ´s ) The Sixth International Conference on Biophysics and Synchrotron Radiation will be held at the Advanced Photon Source, Argonne National Laboratory, from August 4-8, 1998. The conference will feature invited talks, extensive poster sessions, juried "Hot Topics" talks by students and postdoctoral fellows, informal "Meet the Experts" sessions, and a tour of the APS. Scientific topics include macromolecular crystallography, with a particular emphasis on the applications of synchrotron-based crystallography to problems in cell biology; X-ray scattering from non-crystalline systems such as solutions, fibers and membranes; spectroscopies ranging from the hard X-ray to the UV and IR regions of the spectrum; microscopies; medical imaging, apparatus and experimental techniques; and the impact of synchrotron radiation on biophysics (past, present and future). These fields have been expanding very briskly, in response both to earlier scientific successes and to the availability of the new, third-generation synchrotron sources at the European Synchrotron Radiation Facility in Grenoble, France, the Advanced Photon Source itself, and the SPring 8 facility under construction in Japan. The expansion is also fueled by strong industrial interest, principally from the pharmaceutical and biotechnology industries. The conference is likely to attract up to 650 participants, ranging from beginning graduate students to Nobel Laureates
Keywords: X ray crystallography, biophysics, electromagnetic radiation, meeting /conference /symposium, particle accelerator
Project start date: 1998-08-15
Project end date: 1999-08-14
1R13RR013462-01 (1998): $12000
TIME-RESOLVED MACROMOLECULAR CRYSTALLOGRAPHY
John Keith Moffat, Professor
Biochemistry And Molecular Biologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 5R01GM036452-09 from National Institute Of General Medical Sciences IRG: BBCB
Abstract: A complete understanding of biological function at the molecular level requires an understanding both of static structure, and of the changes in structure that occur during processes such as enzyme catalysis, ligand binding and release, photocycling, and protein unfolding/refolding reactions. These changes in structure typically occur very rapidly under physiological conditions; the lifetimes of functionally crucial, structural intermediates typically lie in the range from seconds to microseconds. Such short-lived, intermediate structures are completely inaccessible to conventional x-ray crystallographic approaches. With the advent of very intense, polychromatic synchrotron x-ray sources, static x-ray diffraction patterns can be recorded from single crystals of strongly scattering proteins in the one to hundreds of milliseconds range, using a sensitive Kodak storage phosphor area detector. Advances now being made in the x-ray source itself, and in focussing optics, will reduce these exposure times by one to two orders of magnitude into the tens of microseconds to one millisecond range. If the detector is moved in its plane during the exposure, the Laue diffraction spots become streaks; and if the structure of the molecules in the crystal lattice changes during the exposure, then the intensities of the Laue diffraction streaks change, and constitute the raw data of a time-resolved x-ray crystallographic experiment. This experiment has three components, reaction initiation, in which a structural perturbation is induced, reaction monitoring and data acquisition, in which the x-ray intensities are measured as a function of time after reaction initiation, and data analysis, in which the time-dependent changes in molecular structure are identified. We propose to apply time-resolved crystallography to three main systems the photocycle of photoactive yellow protein that appears to be a simple bacterial photosensor; the photolysis of carboxymyoglobin at low temperature; and the early stages of protein unfolding in the crystal lattice, induced by a temperature jump. The laser techniques necessary for reaction initiation and optical monitoring of the crystals will be further developed. Data analysis strategies aimed at revealing the structures of individual intermediates, rather than just their time-dependent mixture, will be explored
Keywords: X ray crystallography, chemical structure, macromolecule, nonclinical biomedical equipment binding protein, biosensor, carboxyhemoglobin, chemical binding, chemical structure function, chromophore, flash photolysis, ligand, lysozyme, molecular dynamics, myoglobin, nonvisual photosensitivity, phosphotransferase, ribonuclease Rhodospirillales, Staphylococcus, laser, spectrometry, temperature
Project start date: 1990-09-01
Project end date: 1994-11-30
5R01GM036452-09 (1994): $191746
Sponsored Links Excellgen http://Excellgen.com
John Keith Moffat
University Of Chicago
Project start date: 1990-09-01
Project end date: 2012-12-30
BioCARS: A Synchrotron Structural Biology Resource
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 3P41RR007707-15S1 from National Center For Research Resources IRG: ZRG1
Project start date: 1992-09-15
Project end date: 2008-07-31
3P41RR007707-15S1 (2007): $999994
TIME-RESOLVED MACROMOLECULAR CRYSTALLOGRAPHY
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5R37GM036452-19 from National Institute Of General Medical Sciences IRG: NSS
Keywords: X ray crystallography, method development, molecular dynamics, photochemistry, protein structure function, time resolved data, carboxyhemoglobin, chemical binding, chemical kinetics, conformation, heme, hemoglobin, mathematical model, myoglobin, nonvisual photoreceptor, pancreatic ribonuclease, photoactivation, photolysis, rhodopsin, structural biology, temperature, thermodynamics, bioimaging /biomedical imaging, laser
Project start date: 1990-09-01
Project end date: 2004-11-30
5R37GM036452-19 (2004): $389840
NORMAL AND HYBRID MODE LAUE TIME-RESOLVED STUDIES OF PAS DOMAINS
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P41RR007707-130116 from National Center For Research Resources IRG: ZRG1
Keywords: biomedical resource, chemical structure function, time resolved data, structural biology
Project start date: 2004-08-01
Project end date: 2005-07-31
STRUCTURAL STUDIES OF THE PAS DOMAIN OF NIFL
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P41RR007707-130122 from National Center For Research Resources IRG: ZRG1
Keywords: biomedical resource, chemical structure function, structural biology
Project start date: 2004-08-01
Project end date: 2005-07-31
STRUCTURE DETERMINATION OF THE REDOX SENSOR NIFL
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P41RR007707-130218 from National Center For Research Resources IRG: ZRG1
Keywords: biomedical resource, chemical structure, oxidation reduction reaction, structural biology
Project start date: 2004-08-01
Project end date: 2005-07-31
STRUCTURAL STUDIES OF A NOVEL BLUE LIGHT PHOTORECEPTOR
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P41RR007707-130233 from National Center For Research Resources IRG: ZRG1
Keywords: biomedical resource, photobiology, protein structure, receptor, structural biology
Project start date: 2004-08-01
Project end date: 2005-07-31
TIME-RESOLVED MACROMOLECULAR CRYSTALLOGRAPHY
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5R37GM036452-18 from National Institute Of General Medical Sciences IRG: NSS
Keywords: X ray crystallography, method development, molecular dynamics, photochemistry, protein structure function, time resolved data, carboxyhemoglobin, chemical binding, chemical kinetics, conformation, heme, hemoglobin, mathematical model, myoglobin, nonvisual photoreceptor, pancreatic ribonuclease, photoactivation, photolysis, rhodopsin, structural biology, temperature, thermodynamics, bioimaging /biomedical imaging, laser
Project start date: 1990-09-01
Project end date: 2004-11-30
5R37GM036452-18 (2003): $382713
5R37GM036452-17 (2002): $373997
5R37GM036452-16 (2001): $345682
Sponsored Links Excellgen http://Excellgen.com
4R37GM036452-15 (2000): $373714
BIOCARS A SYNCHROTRON STRUCTURAL BIOLOGY RESOURCE
John Keith Moffat, Professor
Biochemistry And Molecular Biologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 2U41RR007707-06 from National Center For Research Resources IRG: ZRG3
Keywords: X ray, X ray crystallography, biomedical equipment resource, biomedical resource, cooperative study, particle accelerator, structural biology
Project start date: 1992-09-15
Project end date: 2002-09-14
2U41RR007707-06 (1997): $2204677
BIOCARS--A SYNCHROTRON STRUCTURAL BIOLOGY RESOURCE
John Keith Moffat, Professor
Biochemistry And Molecular Biologyuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 3U41RR007707-05S1 from National Center For Research Resources IRG: SSS
Project start date: 1992-09-15
Project end date: 1997-09-14
3U41RR007707-05S1 (1997): $1148
CORE RESEARCH--BEAMLINE CONSTRUCTION
John Keith Moffat, Professor
University Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 2U41RR007707-069005 from National Center For Research Resources
Keywords: X ray, biomedical equipment development, biomedical equipment purchase, magnetic field, particle accelerator X ray crystallography, light scattering, molecular biology, optics, time resolved data, virus morphology
DESIGN OF XRAY BEAMLINES FOR STRUCTURAL BIO: HIV NEF PROTEINS STRUCT
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 2U41RR007707-060002 from National Center For Research Resources
Keywords: AIDS, biomedical equipment development, biomedical resource, building /facility design /renovation, communicable disease, im2U41RR007707-069002
Project start date: 1997-09-30
Project end date: 1998-09-14
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 2U41RR007707-069002 from National Center For Research Resources
Keywords: X ray, X ray crystallography, biomedical equipment development, radiation detector, method development, particle accelerator, time resolved data
TIME RESOLVED MACROMOLECULAR CRYSTALLOGRAPHY
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5R37GM036452-11 from National Institute Of General Medical Sciences IRG: BBCB
Project start date: 1990-09-01
Project end date: 1999-11-30
5R37GM036452-11 (1996): $277202
BIOCARS--A SYNCHROTRON STRUCTURAL BIOLOGY RESOURCE
John Keith Moffat, Professor
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5U41RR007707-05 from National Center For Research Resources IRG: SSS
Project start date: 1992-09-15
Project end date: 1997-09-14
5U41RR007707-05 (1996): $1375320
3U41RR007707-01S1 (1993): $1208