Karl Franzens University Graz
Graz University of Technology
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Karl Franzens University Graz | Graz University of Technology | |
Cryo-Electron Microscopy and Tomography: The past, the present and the future The structural elucidation of isolated macromolecules with cryo-EM (i. e. ex situ) has been awarded with the nobel prize in chemistry in 2017 and can be seen nowadays as an established method. Although cryo-EM is not yet one of the high-throughput methods, the requirements in terms of user experience and measurement time are becoming ever less demanding. Equivalent to the beamlines in protein crystallography the first cryo-EM facilities are being build up and put into operation. Of course there is still room for further improvements in cryo-EM. The latest development of the Volta phase plate is a good example of this. The clear phase contrast improves the selection and classification of the individual particles and thus also enables the structural determination of very small proteins that were previously inaccessible. However, proteins have their biological function in the complex environment of the cell and interact with other macromolecules. The exciting potential of cryo-EM therefore lies in cryo-electron tomography, the three-dimensional analysis of biological structures in the undisturbed cellular context (i. e. in situ). This method closes the gap between molecular and cytological structural research. After a a brief look back this lecture will present our recent work in the field of cryo-electron tomography and in situ structural biology and highlights technological developments, limitations and their opportunities. Furthermore, it will give a prospective towards obtaining structural insights from an in situ context, possibly at atomic resolution. |