Electron cryomicroscopy: Difference between revisions
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==Density Maps== | ==Density Maps== | ||
<table align="right"><tr><td> | <table align="right" class="wikitable"><tr><td> | ||
[[Image:6nef-em-map-hec503-his169-360.gif]] | [[Image:6nef-em-map-hec503-his169-360.gif]] | ||
</td></tr><tr><td> | </td></tr><tr><td> | ||
Density map for heme and two histidines in the 3.4 Å cryo-EM structure of a cytochrome ([[6nef]]). | Density map for heme and two histidines in the 3.4<br>Å cryo-EM structure of a cytochrome ([[6nef]]). | ||
</td></tr></table> | </td></tr></table> | ||
The result of a cryo-EM experiment is a density map. Just as for [[X-ray diffraction]], it is then necessary to fit an atomic model optimally into the map<ref name="primer" />. | The result of a cryo-EM experiment is a density map. Just as for [[X-ray diffraction]], it is then necessary to fit an atomic model optimally into the map<ref name="primer" />. | ||
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Electrons are diffracted by the charges in the sample, in contrast to X-rays that are diffracted by electron density, producing [[electron density maps]]. Consequently, EM maps may be termed "electron potential maps"<ref name="rosenthal2019" />, "Coulomb potential maps"<ref name="marques2019">PMID: 31400843</ref>, or "electric potential maps"<ref name="wang2016">PMID: 27706888</ref>. Electron densities are all positive, while electron potential maps can be positive or negative<ref name="wang2016" /><ref name="marques2019" />. Wang (2017)<ref name="wang2017">PMID: 28543856</ref> provided a way to convert electron potential maps to <i>charge density maps</i> (using [[Chimera]]), where densities have better resolution and better reflect the positions of atomic nuclei. | Electrons are diffracted by the charges in the sample, in contrast to X-rays that are diffracted by electron density, producing [[electron density maps]]. Consequently, EM maps may be termed "electron potential maps"<ref name="rosenthal2019" />, "Coulomb potential maps"<ref name="marques2019">PMID: 31400843</ref>, or "electric potential maps"<ref name="wang2016">PMID: 27706888</ref>. Electron densities are all positive, while electron potential maps can be positive or negative<ref name="wang2016" /><ref name="marques2019" />. Wang (2017)<ref name="wang2017">PMID: 28543856</ref> provided a way to convert electron potential maps to <i>charge density maps</i> (using [[Chimera]]), where densities have better resolution and better reflect the positions of atomic nuclei. | ||
===Visualizing EM Maps=== | |||
[http://firstglance.jmol.org FirstGlance in Jmol] makes it easy to visualize EM density maps, as shown in the example at right. Load your [[PDB ID]] (or [[6nef]]). Then, use '''Find..''' (in the [https://proteopedia.org/wiki/fgij/where.htm#focusbox Focus Box]) to locate the residues of interest (or "503,169,360" for 6nef -- In 6nef, these happen to be unambiguous sequence numbers for HEC503, HIS169, and HIS360.) Click on "EM Density Map". After adjusting the view as you wish, click "Save Image or Animation for Powerpoint". An example of an animation saved from FirstGlance is above in this page. | |||
{{Template:PDBMapViewers}} | |||
===The Electron Microscopy Data Bank=== | ===The Electron Microscopy Data Bank=== | ||
The [[Electron Microscopy Data Bank]] (EMDB) archives EM density maps. Only "all features" maps are deposited, as cryo-EM analysis has no equivalent to the [[Electron_density_maps#Fo-Fc_Difference_Map|difference map]] of X-ray diffraction. | The [[Electron Microscopy Data Bank]] (EMDB) archives EM density maps. Only "all features" maps are deposited, as '''cryo-EM analysis has no equivalent to the [[Electron_density_maps#Fo-Fc_Difference_Map|difference map]]''' of X-ray diffraction. | ||
==Temperatures (B factors)== | ==Temperatures (B factors)== | ||