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Whether solid, liquid, or gas, a substance's state depends on the order and arrangement of its particles (atoms, molecules, or ions). Particles in the solid pack closely together, generally in a pattern. The particles vibrate about their fixed positions but do not move or squeeze past their neighbors. In liquids, although the particles are closely spaced, they are randomly arranged. The position of the particles are not fixed—that is, they are free to move past their neighbors to...
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The universe is composed of matter in different forms, and all forms of matter contain energy.  The different forms of energy on Earth originate from the Sun — the ultimate energy source. Plants capture light energy from the Sun, and, via the process of photosynthesis, convert it into chemical energy. This stored energy from plants can be harnessed in many ways. For example, eating plant products as food provides energy for our body to function, and burning wood or coal (fossilized...
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Some solids can transition directly into the gaseous state, bypassing the liquid state, via a process known as sublimation. At room temperature and standard pressure, a piece of dry ice (solid CO2) sublimes, appearing to gradually disappear without ever forming any liquid. Snow and ice sublimate at temperatures below the melting point of water, a slow process that may be accelerated by winds and the reduced atmospheric pressures at high altitudes. When solid iodine is warmed, the solid sublimes...
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Free energy—abbreviated as G for the scientist Gibbs who discovered it—is a measurement of useful energy that can be extracted from a reaction to do work. It is the energy in a chemical reaction that is available after entropy is accounted for. Reactions that take in energy are considered endergonic and reactions that release energy are exergonic. Plants carry out endergonic reactions by taking in sunlight and carbon dioxide to produce glucose and oxygen. Animals, in turn, break...
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Heating a crystalline solid increases the average energy of its atoms, molecules, or ions, and the solid gets hotter. At some point, the added energy becomes large enough to partially overcome the forces holding the molecules or ions of the solid in their fixed positions, and the solid begins the process of transitioning to the liquid state or melting. At this point, the temperature of the solid stops rising, despite the continual input of heat, and it remains constant until all of the solid is...
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Preparation and Delivery of Protein Microcrystals in Lipidic Cubic Phase for Serial Femtosecond Crystallography
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Synchrotron microcrystal native-SAD phasing at a low energy.

Gongrui Guo1,2, Ping Zhu1, Martin R Fuchs2

  • 1Biology Department, Brookhaven National Laboratory, Upton, NY 11973, USA.

Iucrj
|July 19, 2019
PubMed
Summary

Determining biomolecule structures using native single-wavelength anomalous diffraction (SAD) is difficult. This study shows low-energy X-rays enable native SAD phasing from microcrystals, advancing structural biology.

Keywords:
S-SADanomalous diffractionlow-energy X-raysmicrocrystalsmicrodiffractionmultiple crystalsnative SADradiation damage

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Area of Science:

  • Structural biology
  • Biophysics
  • Crystallography

Background:

  • Native single-wavelength anomalous diffraction (SAD) is challenging due to weak anomalous scattering from native elements like sulfur and phosphorus.
  • Low X-ray energies near elemental K-edges enhance anomalous scattering, offering potential for routine native SAD.
  • Applying native-SAD to microcrystals (<10 µm) is particularly difficult due to sample handling and data collection challenges.

Purpose of the Study:

  • To demonstrate the feasibility of native SAD phasing from microcrystals using synchrotron microdiffraction.
  • To establish a robust method for low-energy native SAD phasing with a reduced number of microcrystals.

Main Methods:

  • Exploitation of anomalous microdiffraction signals at 5 keV X-ray energy.
  • Utilization of polyimide wellmounts for microcrystal sample preparation.
  • Application of an iterative crystal and frame-rejection method for data analysis.

Main Results:

  • Successful microcrystal native-SAD phasing was achieved using approximately 1,200 crystals.
  • This method overcomes limitations previously requiring thousands of microcrystals, as seen with X-ray free-electron lasers.
  • Demonstrated the utility of low-energy native-SAD at synchrotron microdiffraction beamlines.

Conclusions:

  • Low-energy native SAD phasing is a viable and robust technique for microcrystals.
  • This approach significantly reduces the number of microcrystals required for structural determination.
  • Advances the application of synchrotron microdiffraction for biomolecular structure analysis.