A conserved histidine in switch-II of EF-G moderates release of inorganic phosphate
2015 (English)In: Scientific Reports, ISSN 2045-2322, Vol. 5, 12970Article in journal (Refereed) Published
Elongation factor G (EF-G), a translational GTPase responsible for tRNA-mRNA translocation possesses a conserved histidine (H91 in Escherichia coli) at the apex of switch-II, which has been implicated in GTPase activation and GTP hydrolysis. While H91A, H91R and H91E mutants showed different degrees of defect in ribosome associated GTP hydrolysis, H91Q behaved like the WT. However, all these mutants, including H91Q, are much more defective in inorganic phosphate (Pi) release, thereby suggesting that H91 facilitates Pi release. In crystal structures of the ribosome bound EF-G center dot GTP a tight coupling between H91 and the gamma-phosphate of GTP can be seen. Following GTP hydrolysis, H91 flips similar to 140 degrees in the opposite direction, probably with Pi still coupled to it. This, we suggest, promotes Pi to detach from GDP and reach the inter-domain space of EF-G, which constitutes an exit path for the Pi. Molecular dynamics simulations are consistent with this hypothesis and demonstrate a vital role of an Mg2+ ion in the process.
Place, publisher, year, edition, pages
2015. Vol. 5, 12970
IdentifiersURN: urn:nbn:se:uu:diva-261233DOI: 10.1038/srep12970ISI: 000359373400001OAI: oai:DiVA.org:uu-261233DiVA: diva2:851842
FunderSwedish Research Council, 2011-6088 2014-4423 2008-6593Knut and Alice Wallenberg Foundation, KAW 2011.0081