Abstract
The NASA mission Perfect Crystals used the microgravity environment on the International Space Station (ISS) to grow crystals of human manganese superoxide dismutase (MnSOD)—an oxidoreductase critical for mitochondrial vitality and human health. The mission’s overarching aim is to perform neutron protein crystallography (NPC) on MnSOD to directly visualize proton positions and derive a chemical understanding of the concerted proton electron transfers performed by the enzyme. Large crystals that are perfect enough to diffract neutrons to sufficient resolution are essential for NPC. This combination, large and perfect, is hard to achieve on Earth due to gravity-induced convective mixing. Capillary counterdiffusion methods were developed that provided a gradient of conditions for crystal growth along with a built-in time delay that prevented premature crystallization before stowage on the ISS. Here, we report a highly successful and versatile crystallization system to grow a plethora of crystals for high-resolution NPC.
| Original language | English |
|---|---|
| Article number | 39 |
| Journal | npj Microgravity |
| Volume | 9 |
| Issue number | 1 |
| DOIs | |
| State | Published - Dec 2023 |
Funding
This research employed GCBs (Granada Crystallization Box) that are no longer manufactured. We thank researchers from all over the world who graciously sent us their GCBs from their inventory to enable the experiments, including Edward Snell (Hauptman Woodward Institute, NY), Joseph Ng (University of Alabama —Huntsville), Alice Dawson (University of Dundee, Scotland), Dominika Borek (University of Southwestern Texas), Janet Newman (CSIRO, Australia), James W. Murray (Imperial College, London)and Dr. Jose Gavira (University of Granada). We also thank Timothy Mueser, April Spinale (CASIS), Edward Snell, and Joseph Ng for useful discussions. We would like to thank Scott Tarry and Michaela Lucas and the NASA Nebraska Space Grant and NASA EPSCoR office at the University of Nebraska—Omaha, and Sridhar Gorti from Marshall Space Flight Center for their support. We thank Uma Verma and Tyler Dorval (Payload Integration Managers from Boeing), Kyle Kesling (Safety Panel Engineer at Johnson Space Center, JSC), Jeppie Compton (National Project Manager with NASA EPSCoR at Kennedy Space Center, KSC), and Willie B. Williams (ISS NASA Research Portfolio Manager at JSC) for managing and making the flights to ISS possible. This research was supported by NIGMS (R01GM145647-01), NASA EPSCoR (NE-80NSSC17M0030 and NE-NNX15AM82A), and NASA space grant fellowships to Jahaun Azadmanesh. The UNMC Structural Biology Core Facility was funded by the Fred and Pamela Buffett NCI Cancer Center Support Grant (P30CA036727). Leighton Coates acknowledges support by the NIH (R01-GM071939). The research at Oak Ridge National Laboratory (ORNL) Spallation Neutron Source was sponsored by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. The Office of Biological and Environmental Research supported research at ORNL Center for Structural Molecular Biology (CSMB) using facilities supported by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. This research employed GCBs (Granada Crystallization Box) that are no longer manufactured. We thank researchers from all over the world who graciously sent us their GCBs from their inventory to enable the experiments, including Edward Snell (Hauptman Woodward Institute, NY), Joseph Ng (University of Alabama —Huntsville), Alice Dawson (University of Dundee, Scotland), Dominika Borek (University of Southwestern Texas), Janet Newman (CSIRO, Australia), James W. Murray (Imperial College, London)and Dr. Jose Gavira (University of Granada). We also thank Timothy Mueser, April Spinale (CASIS), Edward Snell, and Joseph Ng for useful discussions. We would like to thank Scott Tarry and Michaela Lucas and the NASA Nebraska Space Grant and NASA EPSCoR office at the University of Nebraska—Omaha, and Sridhar Gorti from Marshall Space Flight Center for their support. We thank Uma Verma and Tyler Dorval (Payload Integration Managers from Boeing), Kyle Kesling (Safety Panel Engineer at Johnson Space Center, JSC), Jeppie Compton (National Project Manager with NASA EPSCoR at Kennedy Space Center, KSC), and Willie B. Williams (ISS NASA Research Portfolio Manager at JSC) for managing and making the flights to ISS possible. This research was supported by NIGMS (R01GM145647-01), NASA EPSCoR (NE-80NSSC17M0030 and NE-NNX15AM82A), and NASA space grant fellowships to Jahaun Azadmanesh. The UNMC Structural Biology Core Facility was funded by the Fred and Pamela Buffett NCI Cancer Center Support Grant (P30CA036727). Leighton Coates acknowledges support by the NIH (R01-GM071939). The research at Oak Ridge National Laboratory (ORNL) Spallation Neutron Source was sponsored by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. The Office of Biological and Environmental Research supported research at ORNL Center for Structural Molecular Biology (CSMB) using facilities supported by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy.
| Funders | Funder number |
|---|---|
| Fred and Pamela Buffett NCI | P30CA036727 |
| Jeppie Compton | |
| NASA EPSCoR | NE-80NSSC17M0030, NE-NNX15AM82A |
| Scientific User Facilities Division | |
| Uma Verma and Tyler Dorval | |
| University of Nebraska | |
| University of Southwestern Texas | |
| National Institutes of Health | R01-GM071939 |
| Boeing | |
| U.S. Department of Energy | |
| National Institute of General Medical Sciences | R01GM145647-01 |
| National Aeronautics and Space Administration | |
| Basic Energy Sciences | |
| Kennedy Space Center | |
| Marshall Space Flight Center | |
| Biological and Environmental Research | |
| Oak Ridge National Laboratory | |
| University of Dundee | |
| University of Alabama | |
| Kansas Soybean Commission | |
| Universidad de Granada |