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Albany 2019: 20th Conversation - Abstracts

category image Albany 2019
Conversation 20
June 11-15 2019
Adenine Press (2019)

MicroED: conception, practice and future opportunities

My laboratory studies the structures of membrane proteins that are important in maintaining homeostasis in the brain. Understanding structure (and hence function) requires scientists to build an atomic resolution map of every atom in the protein of interest, that is, an atomic structural model of the protein of interest captured in various functional states. In 2013 we unveiled the method Microcrystal Electron Diffraction (MicroED) and demonstrated that it is feasible to determine high-resolution protein structures by electron crystallography of three-dimensional crystals in an electron cryo-microscope (CryoEM) (Shi 2013; Nannenga 2014). The CryoEM is used in diffraction mode for structural analysis of proteins of interest using vanishingly small crystals. The crystals are often a billion times smaller in volume than what is normally used for other structural biology methods like x-ray crystallography. In this seminar I will describe the basics of this method, from concept to data collection, analysis and structure determination, and illustrate how samples that were previously unattainable can now be studied by MicroED. I will conclude by highlighting how this new method is helping us understand major brain diseases like Parkinson’s disease (Rodriguez 2015); helping us discover and design new drugs; shedding new light on chemical synthesis and small molecule chemistry; and showing us unprecedented level of details with sub atomic resolutions.

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This research has been supported by funds from the Howard Hughes Medical Institute.

References

    Shi D., Nannenga B., Iadanza MG. and Gonen T (2013). MicroED – Three dimensional electron crystallography of protein microcrystals. eLife – 2:e01345: 1 - 17.

    Nannenga B., Shi D., Leslie AGW and Gonen T (2014). Continuous rotation structure determination by MicroED. Nature Methods 11 (9) : 927 - 930.

    Rodriguez A.J., Ivanova M., Sawaya MR., Cascio D., Reyes F., Shi D., Sangwan S., Guenther EL., Johnson L, Zhang M., Jiang L., Arbing M., Nannega B., Hattne J., Whitelegge J., Brewster AS., Messerschmidt M., Boutet S., Sauter NK., Gonen T and Eisenberg D (2015) The toxic core of α-synuclein of Parkinson’s disease: structure from invisible crystals. Nature 525 (7570): 486 - 490.

Tamir Gonen

Howard Hughes Medical Institute
University of California, Los Angeles
Los Angeles, CA 90095

Ph: (310) 825 2716
Fax: (571)291 6449
Email: tgonen@ucla.edu