Electron cryomicroscopy: Difference between revisions
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Single-particle '''electron cryomicroscopy''' ('''cryo-EM''') has become an important method for determining macromolecular structures. It is the basis for the [[Nobel Prizes for 3D Molecular Structure#2010-2019|2017 Nobel Prize in Chemistry]]. Although resolution is usually poorer than that obtained by [[X-ray crystallography]], cryo-EM has the great advantage of not requiring crystallization<ref>Obtaining highly-ordered crystals is perhaps the major obstacle to determination of structure by X-ray diffraction. Less than half of cloned, expressed, purified proteins are sufficiently soluble for structure determination. Of these, diffraction-quality crystals are obtained for only about one in five. See [http://www.umass.edu/molvis/workshop/allstruc/xsuccess.htm Structural Genomics Progress, 2011].</ref>. Cryo-EM is particularly suited to determination of the structures of large complexes containing multiple proteins or nucleic acids, often the most difficult to crystallize. | Single-particle '''electron cryomicroscopy''' ('''cryo-EM''') has become an important method for determining macromolecular structures. It is the basis for the [[Nobel Prizes for 3D Molecular Structure#2010-2019|2017 Nobel Prize in Chemistry]]. Although resolution is usually poorer than that obtained by [[X-ray crystallography]], cryo-EM has the great advantage of not requiring crystallization<ref>Obtaining highly-ordered crystals is perhaps the major obstacle to determination of structure by X-ray diffraction. Less than half of cloned, expressed, purified proteins are sufficiently soluble for structure determination. Of these, diffraction-quality crystals are obtained for only about one in five. See [http://www.umass.edu/molvis/workshop/allstruc/xsuccess.htm Structural Genomics Progress, 2011].</ref>. Cryo-EM is particularly suited to determination of the structures of large complexes containing multiple proteins or nucleic acids, often the most difficult to crystallize. | ||
For a quick overview of the method, see [ | For a quick overview of the method, see [http://www.youtube.com/watch?v=BJKkC0W-6Qk this silent 3 min video] by [http://www.lander-lab.com/ Gabe Lander]. For a historical view of the development of modern cryo-EM, see this [http://www.nobelprize.org/uploads/2018/06/advanced-chemistryprize2017.pdf scientific background] for the 2017 Nobel prize. For a more detailed discussion, see this [http://grigoriefflab.janelia.org/sites/default/files/Cheng_Cell2015.pdf primer]<ref>DOI:10.1016/j.cell.2015.03.050</ref> | ||
==Docking crystal structures into cryo-EM maps== | ==Docking crystal structures into cryo-EM maps== | ||