Robert E. Blankenship - US grants

9451116 Bieber The major goals of the project are to develop and perfect a number of NMR and molecular modeling experiments that are suitable for integration into the undergraduate chemistry curriculum. An NMR and molecular modeling software will be purchased. Several excellent NMR experiments have been designed and tested by faculty for the biophysical, inorganic, organic and physical chemistry laboratories. and more than a dozen undergraduate students are engaged in research projects that use NMR. Funding the proposed project will allow faculty to develop additional NMR experiments for undergraduate courses, will expose all majors and many non- majors to NMR, and will permit students to apply powerful molecular modelling tools to chemical problems. The project will make it feasible for other institutions which lack NMR instrumentation to carry out NMR data analyses and molecular modelling studies at modest cost, because NMR data, processing macros and documentation will be available from Arizona State University through electronic communication or in tape formats. By using electronic networks, the impact of the project will extend well beyond undergraduate instruction at Arizona State University.

DNA sequence determinations are of fundamental importance for studies in many areas of the biosciences. including functional biochemistry, cell and molecular biology, population genetics, and molecular systematics. The purpose of this proposal is to streamline DNA sequencing at Arizona State University by seeking funds to establish a DNA sequencing facility. The major piece of equipment for this facility will be an automated DNA sequencer, which currently is not available at our University and which is requested in this proposal. About 20 groups at Arizona State University routinely use DNA sequencing in their research, and a large number of graduate students, undergraduates, postdoctoral fellows, staff, and faculty will benefit from the establishment of a DNA sequencing facility with an automated sequencer. The sequencer requested utilizes an infrared laser diode to excite fluorophor-labeled DNA that is size-separated by gel electrophoresis. Fluorescence from the fluorophor is detected, and simultaneous scanning of four sequencing lanes over time can provide a DNA sequence of 800 nucleotides or more with over 99% accuracy in a highly cost-effective manner. With the equipment requested, 22 samples can be sequenced simultaneously. Apart from providing a 30% cost-share in purchasing this equipment, the University also will create a staff-level position for an individual to run the DNA sequencing facility and will bear most of the personnel cost involved. A Ph.D.-level research scientist (already on staff) will supervise the facility. Nominal user fees (about $5 per sequence) will be charged to cover the cost of chemicals, the DNA sequencer service contract, and a small part (about 20%) of the technician's salary. This facility will provide a highly effective and economic mechanism to obtain and analyze DNA sequence information. An additional benefit is that DNA sequencing becomes accessible to a large number of graduate students who work in research areas (for examp le, systematics and population biology) that would benefit from DNA analysis, but for whom DNA sequencing facilities currently are not readily available. Also, many undergraduate projects, which usually preclude use of radioactivity, could include DNA sequencing. Another significant advantage of acquisition of an automated DNA sequencer is that when DNA sequencing becomes more efficient, students will be able to gather and analyze sequence information more expediently, leaving more room for emphasis on training in other important areas.

The Photosynthesis Center at Arizona State University has developed a major research program investigating fast energy and electron transfer during solar energy conversion in both natural and artificial systems. Ultrafast optical laser spectroscopy is a key tool in this research effort. Six groups, representing about 40 faculty, postdocs and students, are actively using an existing femtosecond time resolution transient absorbance spectrometer to study bacterial, plant and synthetic antenna and reaction center complexes. This research includes designing and synthesizing molecular devices which mimic the fast rates and high yields of the natural systems but are much simpler chemically and may have applications as molecular switches and logic gates. Also, investigation of the structurally well defined reaction centers of the purple nonsulfur bacteria is leading towards an understanding of the photophysics of the initial solar energy conversion event and towards an understanding of the role played by the protein in modifying the thermodynamic and kinetic parameters of electron transfer. Research is also underway into the light harvesting and energy trapping reactions of the Photosystem I reaction center of higher plants as well as in a related photosynthetic reaction center from Heliobacillus mobilis. A great deal of progress has been made with the present instrument, but as the experiments performed have become more and more sophisticated, limitations have been reached in available time resolution, sensitivity, noise suppression, photon excitation densities, available excitation and probe wavelengths, capabilities to perform multiple excitation pulse experiments, and simply in the amount of time available for each group on the apparatus. The purchase of a new instrument is proposed with greatly expanded capabilities. Unlike the present instrument which uses an actively mode-locked Nd:YAG laser synchronously pumping a dye laser as the source of ultrafast pu lses, the new system will employ a self-mode-locked titanium sapphire laser pumped by a CW argon ion laser. The pulses from this source will be amplified in a solid-state regenerative amplifier to the millijoule level at kilohertz repetition rates and used to generate up to two excitation pulses at any wavelengths between 400 nm and 2.4 rnicrons with pulse durations of roughly 50 fs. In addition, part of the regeneratively amplified pulse will be used to generate a white light continuum, allowing simultaneous absorbance change measurements at many different wavelengths in a single experiment. By using optical gating in a nonlinear crystal, the instrument will also have the capability of performing femtosecond time resolution emission decay studies. Purchase of the proposed femtosecond system will open up a number of new research avenues for photosynthesis investigators at ASU. New excitation wavelengths will allow more specific preparation of molecular excited states Multiple excitation pulses at higher photon densities will allow the creation of specific intermediate states with one pulse whose photochemistry can then be investigated with a second excitation event. The higher time resolution of the system will both help in the investigation of processes previously detected but not resolved on the 100 fs timescale and also open up a window into the molecular vibrations of the system coupled to the absorbance transitions and photochemistry. Higher signal-to-noise absorbance change data will allow more thorough investigation of natural photosynthetic systems with large numbers of pigments whose absorbance masks the small absorbance changes associated with the primary photochemistry of solar energy conversion.

9418415 Blankenship Experiments are proposed to extend the current understanding of the energy trapping processes, primary photochemistry and early secondary electron transfer reactions in photosynthetic reaction centers that contain FeS centers as early electron acceptors. The particular systems that will be studied included reaction centers from the primitive anoxygenic organisms known as heliobacteria as well as Photosystem I of oxygenic photosynthetic organisms. Specific proposed experiments include low-temperature picosecond transient absorbance spectroscopic studies of energy transfer and trapping in both heliobacteria and Photosystem I, picosecond transient absorbance measurement in the blue spectral region in both systems, and investigation of secondary electron transfer reactions in heliobacteria monitored using EPR and millisecond timescale transient absorbance spectroscopy. These experiments will give new insights into the energy trapping process in reaction centers that contain integral core antenna pigments and will help to identify and characterize some of the early electron acceptors in these systems, including chlorophylls, quinones and FeS centers. %%% This project will increase our understanding of how plants and other photosynthetic organisms convert the energy of sunlight into chemical energy. The process of photosynthesis supplies all of our food and most of our energy needs. Many aspects of how photosynthesis works are not understood at a deep level. This project is designed to give detailed knowledge of the chemical mechanism of this essential biological process. ***

9727607 Blankenship The objective of this research is to extend the current understanding of the energy trapping processes, primary photochemistry and early secondary electron transfer reactions in photosynthetic reaction centers that contain FeS centers as early electron acceptors. The particular systems that will be studied include reaction centers from the primitive anoxygenic organisms known as heliobacteria as well as Photosystem I of oxygenic photosynthetic organisms. One of the overall goals of the work is to correlate some of the pigments observed in spectroscopic studies with pigments observed in the recent X-ray structure of Photosystem I. Specific proposed experiments include, picosecond transient absorbance measurements in the blue spectral region in Photosystem I, studies on Photosystem I mutants in the quinone-binding region and the P700 binding region, dichroism experiments on Photosystem I and investigation of secondary electron transfer reactions in heliobacteria monitored using EPR and picosecond timescale transient absorbance spectroscopy. These experiments will give new insights into the energy trapping process in reaction centers that contain integral core antenna pigments and will help to identify and characterize some of the early electron acceptors in these systems, including chlorophylls, quinones and FeS centers. The goal of this work is to understand how plants and other photosynthetic organisms convert the energy of sunlight into chemical energy. This project is designed to give detailed knowledge of the chemical mechanism of energy storage in a class of photosynthetic complexes that is not well understood. Advances in understanding the basic scientific principles at work in this essential biological process may ultimately lead to the development of artificial energy storage systems. The artificial systems may decrease our dependence on fossil fuels and reduce emission of pullutants and greenhouse gases. Another possible benefit is the use of this knowledg e in the development of improved plants that will help meet our food and fiber needs. ***

Abstract # 0070356 Integrated AFM-optical microscope (BioScope) for molecular and cell biology
PI: Y. L. Lyubchenko, Arizona State University.

AFM is a novel technique that offers unique advantages with the potential for very high resolution imaging of macromolecules, their complexes and cells in the absence of stains, shadows, and labels. AFM can be performed in air at ambient conditions or in aqueous solutions the latter being particularly important for resolving fully hydrated structures. Recently, instruments which integrate AFM technology with an optical light microscope has permitted cells, microorganisms macromolecular complexes to be imaged with a resolution much greater than the ~200 nm resolution of the best optical microscopes.
A BioScope AFM, an integrated Atomic Force Microscope/Optical Microscope will be used to study the structure of living cells and macromolecular assemblies and to follow biochemical processes in living as well as fixed cells, tissues, and developing organisms. This is one of the most powerful instruments used in cellular and structural biology and material science today. Specifically, this integrated optical/AFM microscope will be able
to locate with the optical microscope the area of interest;
to obtain high resolution topographic images of cells, their components and macromolecular complexes;
to combine optical and topographic characteristics of the samples;
to follow the dynamics of living cells and their components.
These new capabilities will allow more detailed structural and functional studies of a wide range of biological systems including a) the molecular structure of DNA during recombination, b) the structure and spectral properties of photosynthetic complexes in bacteria, c) surface reorganization of sperm just before fertilization of amphibian and mammalian eggs, d) the surface structure of pathogenic fungi, e) the mechanisms of viral interactions with cell surfaces, and f) surface characterization of biomimetic materials used in artificial organs and tissues. In each case, this new imaging technique is expected to provide new insights into how cells interact with both biological and nonbiological surfaces. The BioScope will be housed in the W. M. Keck Bioimaging Laboratory at Arizona State University and will be accessible to any interested party at the University, including faculty, postdoctoral researchers as well as graduate and undergraduate students.

The objective of this proposal is to assess the feasibility of exploiting the unique optical properties of sub-microscopic photosynthetic structures known as chlorosomes for potential use in novel light-converting
and light-detecting device applications. In natural form, chlorosomes serve as light-collecting antennas in some photosynthetic microorganisms which efficiently convert this energy into a form useful for the microorganism. In applied form, we will attempt to marry engineering and biology to take advantage of their unique molecular architecture and their efficient energy transfer processes for use in novel 'biohybrid' device applications where enhanced light-detection, and energy conversion and storage is desired.
In this project, the photosynthesis team proposes to isolate and characterize chlorosome sub-units, in particular their ability to self-assemble, a property that may serve as the basis of novel enabling
nanotechnologies that allow the design and fabrication of molecular-based optical materials, devices and systems. The bioengineering group proposes to assess the feasibility of designing an optical biomolecular hybrid device that incorporates chlorosome assemblies interfaced in programmed ways to selected light detectors and transducers to demonstrate the potential for enhanced device performance from using biohybrid approaches. The potential impact of the project is threefold. It can (a) contribute to the fundamental knowledge of the complex process of photosynthesis, (b) potentially lead to novel devices and systems that have a number of practical applications in areas such as microelectronics, biotechnology, medicine, and (c) further the NSF goal of integrating research and education, particularly across science and engineering
disciplinary boundaries.

Photosynthetic organisms use a twofold strategy to capture and store solar energy. An antenna system first collects light and then transfers the energy to an electron transfer system that converts electronic excitation into redox energy that can be used for cell growth. In most photosynthetic systems, the antenna and electron transfer functions are located on separable complexes and can be studied independently. However, in Photosystem I of oxygenic photosynthetic organisms, the antenna and electron transfer functions are fused into a single large complex that serves both functions. Experiments are proposed to improve our current fragmentary understanding of the energy trapping processes, primary photochemistry and early secondary electron transfer reactions in this important class of photosynthetic reaction centers. Overall goals of the work are to understand the coupling of the antenna system to the electron transfer system and the pathway of the early electron transfer processes.

Specific experiments in this project include: 1) Studies on Photosystem I mutants in the quinone and pigment binding regions, 2) Separation of the Photosystem I reaction center complex into antenna and electron transfer domains using protein engineering and 3) Analysis of Photosystem I energy trapping and electron transfer in a newly-discovered chlorophyll d-containing organism, Acaryochloris marina.

This project will increase our understanding of how plants and other photosynthetic organisms convert the energy of sunlight into chemical energy. The process of photosynthesis supplies all of our food and most of our energy needs. This project is designed to give detailed knowledge of the chemical mechanism of this essential biological process.

A grant has been awarded to Arizona State University under the direction of Dr. Thomas Dowling for the acquisition of a 48-capillary DNA sequencer/genetic analyzer, and an HPLC genotyping and DNA fragment purification system. This instrumentation is essential for efficient completion of the diversity of projects being conducted at ASU, including molecular, evolutionary, ecological, genomic, and bioinformatic studies. Acquisition of this equipment is necessary to handle both increasing demands (number of samples) by existing and new faculty, and will permit investigators to address questions not currently approachable with the existing systems. Research areas that will utilize the instrumentation include: (1) molecular genetics and the evolution of photosynthesis, (2) molecular systematics and evolution of a diversity of organisms (e.g., bacteria, fungi, plants, animals), (3) genetics and management of endangered species, (4) introgressive hybridization and evolutionary genomics, and (5) population biology. The equipment will become part of the DNA sequencing facility at the university. This facility provides support to several educational programs, such as the NSF REU and UMEB programs, which introduce students of diverse backgrounds to careers in science. Central to training these students is exposure to the newest techniques that allow us to obtain previously unobtainable answers. Therefore, addition of this equipment will allow us to better serve the undergraduates and graduates working in our laboratories by exposing them to state of the art technology and techniques.

Complete genome sequencing will be done for four photosynthetic bacteria. Genome sequences of these organisms will fill large gaps in the available genomic data for photosynthetic organisms and will help to understand the origin and early evolution of photosynthesis. In addition, the data that will be obtained will have agricultural applications and environmental importance for understanding global photosynthetic productivity. Photosynthesis has a deep evolutionary connection to nitrogen fixation and many photosynthetic bacteria are capable of nitrogen fixation. Our ability to understand these complex evolutionary relationships and metabolic processes is limited by a lack of data. This genome project will fill major gaps in the evolutionary picture of photosynthesis and its relation to other biological processes. The organisms that will be sequenced in this project are Heliobacterium modesticaldum, Roseobacter denitrificans, Rhodocista centenaria and Acaryochloris marina. The project will actively engage a large number of bioinformatics students in the annotation efforts. Bioinformatics graduate students will use the raw genome data as part of "real world" class exercises to identify metabolic pathways or families of transport proteins. The project team includes experts on each organism, including Robert Blankenship, Michael Madigan, Thomas Beatty, Carl Bauer, Mamoro Mimuro and Hideaki Miyashita and a highly experienced sequencing center, directed by Jeffrey Touchman. The team will also partner with the public science museum at the Arizona Science Center to develop public displays and teacher training materials aimed at communicating the excitement and benefits of microbial genomics to the general public and to school children, respectively.

Chemistry (12) This project aims to introduce mass spectrometry (MS) into all levels of the undergraduate chemistry and biochemistry curriculum at Arizona State University. A major objective of the project is to use MS as a means for developing "molecular thinking" in students. Acquisition of Gas Chromatography/Mass Spectrometry (GC/MS) and Matrix-Assisted Laser Desorption Ionization - Time of Flight (MALDI-TOF) mass spectrometers are enabling students to perform experiments using mass spectrometry in course-related laboratory work and independent student research. A number of experiments, with increasing complexity, are being adapted from the current scientific literature for instructional purposes at various levels of the curriculum including general chemistry, organic chemistry, instrumental analysis and biochemistry. Assessment of the outcomes of the project is being accomplished through a variety of methods including ongoing evaluation of students, TAs and faculty, as well as exit interviews and tests designed to measure how well the methods help students learn key mass spectrometry and chemistry concepts.

This SGER project is concerned with the characterization of a putative new class of enzymes evolutionarily related to nitrogenase. These proteins have been discovered using genomic methods in a number of organisms, including several that are not diazotrophic. The proteins are evolutionarily related to nitrogenase but almost certainly catalyze other, as yet unknown, chemical reactions. These proteins may be the direct descendents of the primitive ancestors of nitrogenase, because of their basal position in the phylogenetic tree and the single type of subunit in the reductase portion of the complex. In this project, genes for these nitrogenase-like proteins (NflH and NflD) will be cloned from Methanococcus jannaschii and will be expressed in E. coli. Using antibodies obtained from these recombinant proteins, native forms of these same proteins from M. jannaschii will be purified and characterized. It is possible that a unique activity will be determined for these proteins, which could imply a role for a pre-nitrogenase homolog on the early Earth. A further understanding of the mechanism of nitrogenase might also be realized, if a similar substrate or activity is found in these homologs.

Broader Impacts: This project will enhance our understanding of the origin and evolution of the nitrogen cycle, which will give important insights into the early evolution of life and the origin of metabolism. This project will involve training of a postdoctoral fellow and will contribute to the development of a scientifically trained workforce.

With support from the Chemistry Research Instrumentation and Facilities: Departmental Multiuser Instrument Acquisition (CRIF-MU) Program, the Department of Chemistry at Arizona State University will acquire an ultrafast, multidimensional fluorescence detection and imaging system. The instrument has time and imaging capabilities. There are four research areas highlighted: photosynthetic and biommimetic systems, directed molecular evolution using synthetic libraries, nanostructures surfaces, and cell and tissue imaging studies.

Measuring time resolved fluorescence affords researchers direct insight into the local environment and dynamics around a probe. Techniques developed tomonitor time resolved fluorescence have been broadly applied to established fields such as photobiology as well as newly emerging areas in materials chemistry.

A new class of integral membrane oxidoreductase complex was recently discovered in bacteria (Yanyushin et al. Biochemistry, 44, 10037-10045, 2005). The complex was purified from the filamentous green anoxygenic phototroph Chloroflexus aurantiacus. The complex consists of seven subunits, the genes for which were identified using trypsin disgestion fingerprinting, mass spectroscopy and the partially completed genome sequence. Three of the subunits appear to be integral membrane proteins based on hydropathy plots. Two of the subunits contain c-type heme cofactors, based on both heme staining and sequence data. Another subunit has sequence homology at the N-terminal end to a molybdopterin enzyme and the C-terminal end to an FeS protein, although the complex does not contain Mo or pterin cofactors, and the primary sequence is missing key elements of their binding sites. The complex has been named the MFIcc complex. Similar although distinct complexes were isolated from phototrophically and aerobically grown cells, and appear to be coded for by distinct genes for at least most of the subunits. The genes that code for the protein subunits of the complexes are arranged in a cluster on the genome and form a putative operon. What appear to be homologous operons for similar complexes were identified in genomes of a number of nonphotosynthetic bacteria. The presence of genes for this complex are anticorrelated with the presence of genes for the cytochrome bc1 complex, suggesting that the new complex may functionally replace the cytochrome bc1 complex. Chloroflexus aurantiacus almost certainly does not contain a bc1 complex based on both biochemical studies and the lack of genes in its genome that code for the protein subunits. This project involves complete biochemical and energetic characterization of this complex. This includes determination of overall oligomeric state and numbers of each subunit by the use of gel filtration and blue-native and SDS gel electrophoresis, coupled with mass spectroscopy and protein cross-linking studies. Additional information about the redox centers and their midpoint potentials will be obtained using redox potentiometry coupled with UV-Vis and EPR spectroscopies. Quantitative determination of the number and types of cofactors will be carried out using a variety of standard techniques.

Broader Impacts
The information obtained on these complexes will provide new insights into novel bacterial bioenergetic mechanisms and the origin and evolution of energy conserving systems. This project will train a graduate student and a postdoctoral fellow in the integrated techniques of biochemistry, molecular biology, biophysical analysis and bioinformatics that are essential parts of modern research in bioenergetic systems. Undergraduate students will also be involved in all aspects of the research.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The chemical reactions leading to long-term energy storage in photosynthetic systems take place within the membrane-bound reaction center complex and an associated group of proteins that make up an electron transport chain. One of the central goals of our research is to identify the molecular parameters responsible for the fact that essentially every photon absorbed by the system leads to stable products. To this end, we do a variety of kinetic, thermodynamic and structural measurements on antenna complexes, reaction centers, electron transport proteins and isolated pigments, using a number of techniques, including ultrafast laser flash photolysis and UV-VIS, fluorescence and electron spin resonance spectroscopies, as well as biochemical and molecular biological analysis. The appearance of photosynthesis and other metabolic processes such as nitrogen fixation had profound effects on the evolution of advanced life on Earth. Our analysis of whole bacterial genomes has revealed that these metabolic processes have complex evolutionary histories, including substantial horizontal gene transfer. We have also used a combination of genomic, molecular evolution techniques and biochemical analysis to identify and characterize previously unknown enzyme complexes with novel activities.

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 high excitation energy transfer efficiency observed in photosynthetic organisms relies on the optimal pigment-protein binding geometry in the individual protein complexes and also on the overall architecture of photosystems. In green sulfur bacteria, the membrane-attached Fenna-Matthews-Olson (FMO) antenna protein functions as a "wire" to connect the large peripheral chlorosome antenna complex with the reaction center (RC), which is embedded in the cytoplasmic membrane. Energy collected by the chlorosome is funneled through the FMO to the RC. Significant effort has been expanded to understand the relationship between structure and function of the individual isolated particles. The question of how the FMO protein interacts with the membrane and the chlorosome in vivo to maintain a specific architecture to ensure the highly efficient energy transfer pathway, however, has not been answered.

Drs. J. Touchman (Arizona State Univ), R. Blankenship (Washinton State Univ) and M. Madigan (Southern Illinois Univ.) are carrying out the genome sequencing and metabolic analysis of five photosynthetic prokaryotes. Organisms for this project were chosen to provide significantly wider coverage of genomes of phototrophic taxa than is now available. The broad goals of the project are to use the genomic data to understand the origin and evolution of photosynthesis and to explore mechanisms of the anoxygenic to oxygenic transition. The organisms being sequenced include species that live at low temperature (psychrophilic), high temperature (thermophilic), high pH (alkaliphilic), in environments subject to periodic drying, and environments high in sulfide. The organisms chosen include two heliobacteria, Heliorestis convoluta and Heliophilum fasciatum; four proteobacteria, Rhodoferax antarcticus, Rhodopila globiformis, Blastochloris viridis, and Thermochromatium tepidum; and one cyanobacterium, Leptolyngbya (a.k.a. Oscillatoria) amphigranulata. The proteobacteria include members from the beta and gamma divisions and one that contains bacteriochlorophyll b as its principal photopigment. The finished, annotated genome sequences are being used to fill large gaps in the available genomic data for photosynthetic prokaryotes. The metabolic capabilities of these organisms are also being analyzed using pathway analysis software tools. Each organism has individual characteristics that justify its inclusion in a genome-sequencing project, including evolutionary relationships, agricultural applications and environmental aspects.
Dr. Touchman is involved in the Be a Biologist program at ASU. All of the investigators are involving undergraduate and high school students in sequence analysis and annotation activities in their labs.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Photosynthesis is a central biological process that produces all the food and much of the energy used by human beings. An enormous amount of work has been done to understand the structural basis of light harvesting and the efficiency of the energy-transfer process. Several bacterial light-harvesting (LH) complexes, such as LH1, LH2 and Fenna-Matthews-Olson protein (FMO) have served as model systems for structural, spectroscopic and theoretical studies.. The recently discovered, thermophilic bacterium, Candidatus Chloracidobacterium (C.) thermophilum, belongs to the phylum Acidobacteria, and is the only chlorophototroph found in the phylum so far. Surprisingly, the photosystem of the aerobic C. thermophilum closely resembles that of the green sulfur bacteria (GSB), which are strict anaerobes. However, The mechanism that enables the C. thermophilum cells to carry out phototrophy under aerobic conditions is not yet known. FMO proteins of green sulfur bacteria (Chlorobiales) have been extensively studied using a wide range of spectroscopic and theoretical approaches. The new FMO protein is investigated by several approaches including native electrospray in a Bruker 12 tesla FTICR mass spectrometer.

With this award from the Major Research Instrumentation Program (MRI) and support from the Chemistry Research Instrumentation Program (CRIF), Professor Liviu Mirica from Washington University and colleagues Robert Blankenship, William Buhro, D. Andre d'Avignon and Sophia Hayes will acquire an EPR spectrometer that would operate at X-band. This instrument will allow research in a variety of fields such as those that provide insight on how biologically relevant species behave when they possess unpaired electrons. In general, an EPR spectrometer yields detailed information on the geometric and electronic structure of molecular and solid state materials. It may also be used to obtain information about the lifetimes of free radicals, short-lived, highly reactive species involved in valuable chemical transformations as well as the initiation of pathological tumor growth. These studies will impact a number of areas, from the synthesis of inorganic and organic molecules to the development of new solid state materials to compounds of magnetic and biological interest. Employing examples inspired from ongoing research, this instrument will be an integral part of research and teaching at the undergraduate and graduate levels at Washington University as well as institutions in the entire St. Louis area.

The award is aimed at enhancing research and education at all levels, especially in areas such as (a) investigating and developing metal-based catalytic systems; (b) characterizing metal-containing biomolecules biological importance; (c) studying biological macromolecular assemblies relevant to solar energy conversion; (d) characterizing nanomaterials; and (e) understanding the electronic characteristics of inorganic compounds.