A Combined Genetic, Biochemical, and Biophysical Analysis of the A1 Phylloquinone Binding Site of Photosystem I from Green Algae

This project has resulted in the increase in our understanding of how proteins interact with and influence the properties of bound cofactors. This information is important for several reasons, including providing essential information for the re-engineering of biological molecules, such as proteins, for either improved function or entirely new ones. In particular, we have found that a molecule, such as the phylloquinone used in Photosystem I (PS1), can be made a stronger electron donor by placing it in a hydrophobic (greasy) environment surrounded by negative charges. In addition, the protein is constrained in its interactions with the phylloqinone, in that it must bind the cofactor tightly, but not in such a way that would stabilize the reduced (natively-charged) version of the molecule. We have used a combination of molecular genetics, in order to make specific mutations in the region of the phylloquinone, and an advanced form of spectroscopy capable of monitoring the transfer of electrons within PS1 using living cells as the material. This approach turned out to produce a significant savings in time and supplies, as it allowed us to focus quickly on the mutants that produced interesting effects, without having to go through laborious purification of the affected proteins. We followed up selected mutants using other spectroscopic techniques in order to gain more specialized information

A Combined Genetic, Biochemical, and Biophysical Analysis of the A1 Phylloquinone Binding Site of Photosystem I from Green Algae

This project has resulted in the increase in our understanding of how proteins interact with and influence the properties of bound cofactors. This information is important for several reasons, including providing essential information for the re-engineering of biological molecules, such as proteins, for either improved function or entirely new ones. In particular, we have found that a molecule, such as the phylloquinone used in Photosystem I (PS1), can be made a stronger electron donor by placing it in a hydrophobic environment surrounded by negative charges. In addition, the protein is constrained in its interactions with the phylloqinone, in that it must bind the cofactor tightly, but not in such a way that would stabilize the reduced (negatively-charged) version of the molecule. We have used a combination of molecular genetics, in order to make specific mutations in the region of the phylloquinone, and an advanced form of spectroscopy capable of monitoring the transfer of electrons within PS1 using living cells as the material. This approach turned out to produce a significant savings in time and supplies, as it allowed us to focus quickly on the mutants that produced interesting effects, without having to go through laborious purification of the affected proteins. We followed up selected mutants using other spectroscopic techniques in order to gain more specialized information. In addition to the main project funded by this work, this grant supported several related side-projects that also increased our understanding about related issues

Initial Technology Assessment for the Large-Aperture UV-Optical-Infrared (LUVOIR) Mission Concept Study

The NASA Astrophysics Division's 30-Year Roadmap prioritized a future large-aperture space telescope operating in the ultra-violet/optical/infrared wavelength regime. The Association of Universities for Research in Astronomy envisioned a similar observatory, the High Definition Space Telescope. And a multi-institution group also studied the Advanced Technology Large Aperture Space Telescope. In all three cases, a broad science case is outlined, combining general astrophysics with the search for biosignatures via direct-imaging and spectroscopic characterization of habitable exoplanets. We present an initial technology assessment that enables such an observatory that is currently being studied for the 2020 Decadal Survey by the Large UV/Optical/Infrared (LUVOIR) surveyor Science and Technology Definition Team. We present here the technology prioritization for the 2016 technology cycle and define the required technology capabilities and current state-of-the-art performance. Current, planned, and recommended technology development efforts are also reported

Functional Modeling Identifies Paralogous Solanesyl-diphosphate Synthases That Assemble the Side Chain of Plastoquinone-9 in Plastids

Background: Plastid isoforms of solanesyl-diphosphate synthase catalyze the elongation of the prenyl side chain of plastoquinone-9. Results: Corresponding mutants display lower levels of plastoquinone-9 and plastochromanol-8 and display intact levels of vitamin E. Conclusion: Plastochromanol-8 originates from a subfraction of non-photoactive plastoquinol-9 and is not essential for seed longevity. Significance: Viable plastoquinone-9 mutants are invaluable tools for understanding plastid metabolism

Initial Technology Assessment for the Large UV-Optical-Infrared (LUVOIR) Mission Concept Study

The NASA Astrophysics Divisions 30-Year Roadmap prioritized a future large-aperture space telescope operating in the ultra-violet-optical-infrared wavelength regime. The Association of Universities for Research in Astronomy envisioned a similar observatory, the High Definition Space Telescope. And a multi-institution group also studied the Advanced Technology Large Aperture Space Telescope. In all three cases, a broad science case is outlined, combining general astrophysics with the search for bio-signatures via direct-imaging and spectroscopic characterization of habitable exo-planets. We present an initial technology assessment that enables such an observatory that is currently being studied for the 2020 Decadal Survey by the Large UV-Optical Infrared (LUVOIR) surveyor Science and Technology Definition Team. We present here the technology prioritization for the 2016 technology cycle and define the required technology capabilities and current state-of-the-art performance. Current, planned, and recommended technology development efforts are also reported

Soluble extract from the nematode Strongyloides stercoralis induces CXCR2 dependent/IL-17 independent neutrophil recruitment.

Neutrophil recruitment via CXCR2 is required for innate and adaptive protective immunity to the larvae of Strongyloides stercoralis in mice. The goal of the present study was to determine the mechanism of CXCR2-mediated neutrophil recruitment to S. stercoralis. Mice deficient in the receptor for IL-17A and IL-17F, upstream mediators of CXCR2 ligand production, were infected with S. stercoralis larvae; there was no difference in larval survival, neutrophil recruitment, or production of CXCR2 ligands compared with wild type mice. In vivo and in vitro stimulation of neutrophils with S. stercoralis soluble extract resulted in significant neutrophil recruitment. In vitro assays demonstrated that the recruitment functioned through both chemokinesis and chemotaxis, was specific for CXCR2, and was a G protein-coupled response involving tyrosine kinase and PI3K. Finally, neutrophil stimulation with S. stercoralis soluble extract induced release of the CXCR2 ligands MIP-2 and KC from neutrophils, thereby potentially enhancing neutrophil recruitment

Signaling through Galphai2 protein is required for recruitment of neutrophils for antibody-mediated elimination of larval Strongyloides stercoralis in mice.

The heterotrimeric guanine nucleotide-binding protein Galphai2 is involved in regulation of immune responses against microbial and nonmicrobial stimuli. Galphai2-/- mice have a selectively impaired IgM response consistent with a disorder in B cell development yet have augmented T cell effector function associated with increased production of IFN-gamma and IL-4. The goal of the present study was to determine if a deficiency in the Galphai2 protein in mice would affect the protective immune response against Strongyloides stercoralis, which is IL-4-, IL-5-, and IgM-dependent. Galphai2-/- and wild-type mice were immunized and challenged with S. stercoralis larvae and analyzed for protective immune responses against infection. Galphai2-/- mice failed to kill the larvae in the challenge infection as compared with wild-type mice despite developing an antigen-specific Th2 response characterized by increased IL-4, IL-5, IgM, and IgG. Transfer of serum collected from immunized Galphai2-/- mice to naïve wild-type mice conferred passive protective immunity against S. stercoralis infection thus confirming the development of a protective antibody response in Galphai2-/- mice. Differential cell analyses and myeloperoxidase assays for quantification of neutrophils showed a significantly reduced recruitment of neutrophils into the microenvironment of the parasites in immunized Galphai2-/- mice. However, cell transfer studies demonstrated that neutrophils from Galphai2-/- mice are competent in killing larvae. These data demonstrate that Galphai2 signaling events are not required for the development of the protective immune responses against S. stercoralis; however, Galphai2 is essential for the recruitment of neutrophils required for host-dependent killing of larvae

Overview of Spirit Microscopic Imager Results

This paper provides an overview of Mars Exploration Rover Spirit Microscopic Imager (MI) operations and the calibration, processing, and analysis of MI data. The focus of this overview is on the last five Earth years (2005-2010) of Spirit's mission in Gusev crater, supplementing the previous overview of the first 450 sols of the Spirit MI investigation. Updates to radiometric calibration using in-flight data and improvements in high-level processing are summarized. Released data products are described, and a table of MI observations, including target/feature names and associated data sets, is appended. The MI observed natural and disturbed exposures of rocks and soils as well as magnets and other rover hardware. These hand-lens-scale observations have provided key constraints on interpretations of the formation and geologic history of features, rocks, and soils examined by Spirit. MI images complement observations by other Spirit instruments, and together show that impact and volcanic processes have dominated the origin and evolution of the rocks in Gusev crater, with aqueous activity indicated by the presence of silica-rich rocks and sulfate-rich soils. The textures of some of the silica-rich rocks are similar to terrestrial hot spring deposits, and observations of subsurface cemented layers indicate recent aqueous mobilization of sulfates in places. Wind action has recently modified soils and abraded many of the rocks imaged by the MI, as observed at other Mars landing sites. Plain Language Summary The Microscopic Imager (MI) on NASA's Spirit rover returned the highest-resolution images of the Martian surface available at the time of the 2004-2010 mission. Designed to survive 90 Mars days (sols) and search for evidence of water in the past, Spirit returned data for 2210 sols, far exceeding all expectations. This paper summarizes the scientific insights gleaned from the thousands of MI images acquired during the last 5years of the mission, supplementing the summary of the first 450 sols of the Spirit MI investigation published previously (Herkenhoff et al., ). Along with data from the other instruments on Spirit, MI images guided the scientific interpretation of the geologic history of the rocks and soils observed in Gusev crater on Mars. We conclude that the geologic history of the area explored by Spirit has been dominated by impacts and volcanism, and that water, perhaps very hot water, was involved in the evolution of some of the rocks and soils. More recently, winds have moved soil particles and abraded rocks, as observed elsewhere on Mars. These results have improved our understanding of Mars' history and informed planning of future missions to Mars.National Aeronautics and Space AdministrationPublic domain articleThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]