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Large Volume Crystal Growth of Superoxide Dismutase Complexes in Microgravity for Neutron Diffraction Studies

NE

University of Nebraska/NASA Johnson Space Center, Human Exploration & Operations Mission Directorate, International Space Station Superoxide dismutases (SODs) are important antioxidant enzymes that protect all living cells against toxic oxygen metabolites, also known as Reactive Oxygen Species (ROS). SODs are the first line of defense to protect organisms against metabolic- and ionizing radiation-induced ROS. Mutations in SOD lead to degenerative diseases such as amyotrophic lateral sclerosis (ALS), diabetes, and cancer. Despite the biological and medical importance of SOD, the enzymatic mechanism is still unknown; precise structural data are needed to understand it. This proposal will study SODs from the model system Escherichia coli as they are easy to produce, stable, and the active sites are identical with human homologs. Bacteria have both Fe and MnSOD. The binding sites of the diatomic substrate and product as well as the source of the protons in the reaction have been studied but their exact identification has not been possible. This detailed information can only be determined by neutron diffraction. Complexes of the Fe and MnSOD including structural intermediates and mutants will be the targets for large volume crystal (1 mm cubed) growth for structure determination by Neutron Macromolecular Crystallography (NMC). SOD crystals large enough for neutron studies were successfully grown by the Borgstahl laboratory on the International Space Station (ISS) in 2001. Innovation and imagination are all that are required to use the Light With NASA’s renewed interest in protein Microscopy Module (LMM) as a crystal growth aboard the ISS we would like laboratory microscope to perform to move forward with these exciting early research aboard the International microgravity crystallization results for SOD. Space Station (ISS). The LMM, used here by Italian astronaut Paolo Existing crystallization facilities, such as the Nespoli, a remotely controllable, Granada Box Facility (GBF) that employs capillary time—the effects of the space counterdiffusion protocols, or the Protein environment on physics and biology. Specimens can be studied without the Crystallization Facility (PCF) that uses vapor need to return the samples to Earth. diffusion methods, will be used to achieve these goals. NMC will be performed with collaborators at Oak Ridge National Laboratory.

Gloria Borgstahl, Ph.D., Science PI, Professor, Eppley Institute, University of Nebraska Medical Center www.nasa.gov/epscor/stimuli

The LMM flight unit features a modified commercial laboratory Leica RXA microscope configured to operate in an automated mode with interaction from the ground support staff. Its core capabilities include a level of containment, white light imaging, fluorescence, confocal microscopy (available in 2016 to 2017), and an imaging capability from a Q-Imaging Retiga 1300 camera. Sridhar Gorti, Ph.D, NASA Technical Monitor, Marshall Space Flight Center NASA EPSCoR Stimuli 2014-15

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EPSCoR Stimuli 2014-15  

NASA Office of Education’s Aerospace Research & Career Development (ARCD) is pleased to release NASA EPSCoR Stimuli, a collection of univers...

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