A conserved P-loop anchor limits the structural dynamics that mediate nucleotide dissociation in EF-Tu

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Date
2015
Authors
Mercier, Evan
Girodat, Dylan
Wieden, Hans-Joachim
Journal Title
Journal ISSN
Volume Title
Publisher
Nature Research
Abstract
The phosphate-binding loop (P-loop) is a conserved sequence motif found in mononucleotide-binding proteins. Little is known about the structural dynamics of this region and its contribution to the observed nucleotide binding properties. Understanding the underlying design principles is of great interest for biomolecular engineering applications. We have used rapid-kinetics measurements in vitro and molecular dynamics (MD) simulations in silico to investigate the relationship between GTP-binding properties and P-loop structural dynamics in the universally conserved Elongation Factor (EF) Tu. Analysis of wild type EF-Tu and variants with substitutions at positions in or adjacent to the P-loop revealed a correlation between P-loop flexibility and the entropy of activation for GTP dissociation. The same variants demonstrate more backbone flexibility in two N-terminal amino acids of the P-loop during force-induced EF-Tu-GTP dissociation in Steered Molecular Dynamics simulations. Amino acids Gly18 and His19 are involved in stabilizing the P-loop backbone via interactions with the adjacent helix C.We propose that these P-loop/helix C interactions function as a conserved P-loop anchoring module and identify the presence of P-loop anchors within several GTPases and ATPases suggesting their evolutionary conservation.
Description
Sherpa Romeo green journal. Open access article. Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License (CC BY-NC-SA 4.0) applies.
Keywords
Phosphate-binding loop , P-loop , Nucleotide , GTP , EF-Tu , Biomolecular engineering , Kinetics , Translation
Citation
Mercier, E., Girodat, D., & Wieden, H.-J. (2015). A conserved P-loop anchor limits the structural dynamics that mediate nucleotide dissociation in EF-Tu. Scientific Report, 5, 7677. https://doi.org/10.1038/srep07677
URI
https://hdl.handle.net/10133/5678
Collections
Wieden, Hans-Joachim