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Resolving the end-point of protein production
November 2016. In a large collaborative project between Goethe University in Frankfurt, EMBL in Heidelberg, and Gene Center in Munich, the final macromolecular RNA-protein complex in protein biosynthesis has been trapped and structurally resolved. The central dogma of life has to do with the translation of genetic information into protein in the ribosome, the factory of all proteins. Central player for translation is the essential and highly dynamic protein ABCE1, that is burning energy in form of ATP to split subunits of the ribosome and free them for the next round of translation. The ‘iron hammer’ head of ABCE1 has been identified to undergo a large rotation and push the subunits of the ribosome apart, in the process of splitting.
The so called post-splitting complex has been trapped by biochemical expertise in Frankfurt using ancient organisms called Archaea. Chemical linking fixed the integrity of the complex before it was chopped into pieces, while preserving original distance information. In Heidelberg, advanced mass-spectrometry, called XL-MS analysis, revealed structural information between the linked pieces at nanometer scale. The reconstituted post-splitting was analyzed by cryo-EM in Munich, where the complex was deep-frozen and rayed by electrons in the cryo-electron microscope. 2D images of the complex were merged and allowed reconstruction of a 3D Model of the post-splitting state with tightly bound ABCE1.
‘We forced the rebellious and aggressive multi-domain ATPase ABCE1 into a new state on the ribosome and condensed the expertise of three institutes into future textbook knowledge’, captures Robert Tampé in a nutshell. In the future, this work will contribute to answer the still open question of how the cycle of translation and life may be closed. More ...
Publication:
Kristin Kiosze-Becker, Alessandro Ori, Milan Gerovac, André Heuer, Elina Nürenberg-Goloub, Umar Jan Rashid, Thomas Becker, Roland Beckmann, Martin Beck & Robert Tampé (2016). Structure of the Ribosome Post-Recycling Complex Probed by Chemical Cross-Linking and Mass Spectrometry. Nature Communications 7:13248, published online 8 November 2016, doi 10.1038/NCOMMS13248, Link ...
Contact:
Robert Tampé, Institute of Biochemistry, Campus Riedberg, Goethe University Frankfurt, Germany, Tel. +49 69 798-29475, tampe@em.uni-frankfurt.de.