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Structural insights into fusidic acid resistance and sensitivity in EF-G

Hansson, Sebastian LU ; Singh, Ranvir LU ; Gudkov, AT; Liljas, Anders LU and Logan, Derek LU (2005) In Journal of Molecular Biology 348(4). p.939-949
Abstract
Fusidic acid (FA) is a steroid antibiotic commonly used against Gram positive bacterial infections. It inhibits protein synthesis by stalling elongation factor G (EF-G) on the ribosome after translocation. A significant number of the mutations conferring strong FA resistance have been mapped at the interfaces between domains G, III and V of EF-G. However, direct information on how such mutations affect the structure has hitherto not been available. Here we present the crystal structures of two mutants of Thermus thermophilus EF-G, G16V and T84A, which exhibit FA hypersensitivity and resistance in vitro, respectively. These mutants also have higher and lower affinity for GTP respectively than wild-type EF-G. The mutations cause significant... (More)
Fusidic acid (FA) is a steroid antibiotic commonly used against Gram positive bacterial infections. It inhibits protein synthesis by stalling elongation factor G (EF-G) on the ribosome after translocation. A significant number of the mutations conferring strong FA resistance have been mapped at the interfaces between domains G, III and V of EF-G. However, direct information on how such mutations affect the structure has hitherto not been available. Here we present the crystal structures of two mutants of Thermus thermophilus EF-G, G16V and T84A, which exhibit FA hypersensitivity and resistance in vitro, respectively. These mutants also have higher and lower affinity for GTP respectively than wild-type EF-G. The mutations cause significant conformational changes in the switch 11 loop that have opposite effects on the position of a key residue, Phe90, which undergoes large conformational changes. This correlates with the importance of Phe90 in FA sensitivity reported in previous studies. These structures substantiate the importance of the domain G/domain III/domain V interfaces as a key component of the FA binding site. The mutations also cause subtle changes in the environment of the "P-loop lysine", Lys25. This led us to examine the conformation of the equivalent residue in all structures of translational GTPases, which revealed that EF-G and eEF2 form a group separate from the others and suggested that the role of Lys25 may be different in the two groups. (c) 2005 Elsevier Ltd. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Molecular Biology
volume
348
issue
4
pages
939 - 949
publisher
Elsevier
external identifiers
  • wos:000228876400012
  • pmid:15843024
  • scopus:17444373920
ISSN
1089-8638
DOI
10.1016/j.jmb.2005.02.066
language
English
LU publication?
yes
id
c3ad27c4-9987-4e1c-aa93-c91c82176150 (old id 152256)
date added to LUP
2007-07-09 15:25:43
date last changed
2017-09-10 04:38:14
@article{c3ad27c4-9987-4e1c-aa93-c91c82176150,
  abstract     = {Fusidic acid (FA) is a steroid antibiotic commonly used against Gram positive bacterial infections. It inhibits protein synthesis by stalling elongation factor G (EF-G) on the ribosome after translocation. A significant number of the mutations conferring strong FA resistance have been mapped at the interfaces between domains G, III and V of EF-G. However, direct information on how such mutations affect the structure has hitherto not been available. Here we present the crystal structures of two mutants of Thermus thermophilus EF-G, G16V and T84A, which exhibit FA hypersensitivity and resistance in vitro, respectively. These mutants also have higher and lower affinity for GTP respectively than wild-type EF-G. The mutations cause significant conformational changes in the switch 11 loop that have opposite effects on the position of a key residue, Phe90, which undergoes large conformational changes. This correlates with the importance of Phe90 in FA sensitivity reported in previous studies. These structures substantiate the importance of the domain G/domain III/domain V interfaces as a key component of the FA binding site. The mutations also cause subtle changes in the environment of the "P-loop lysine", Lys25. This led us to examine the conformation of the equivalent residue in all structures of translational GTPases, which revealed that EF-G and eEF2 form a group separate from the others and suggested that the role of Lys25 may be different in the two groups. (c) 2005 Elsevier Ltd. All rights reserved.},
  author       = {Hansson, Sebastian and Singh, Ranvir and Gudkov, AT and Liljas, Anders and Logan, Derek},
  issn         = {1089-8638},
  language     = {eng},
  number       = {4},
  pages        = {939--949},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {Structural insights into fusidic acid resistance and sensitivity in EF-G},
  url          = {http://dx.doi.org/10.1016/j.jmb.2005.02.066},
  volume       = {348},
  year         = {2005},
}