About this item:

46 Views | 31 Downloads

Author Notes:

Correspondence: eortlun@emory.edu

Conceptualization, C.D.O., E.A.G., E.A.O.; Methodology, C.D.O., M.F.C., E.A.O.; Investigation, C.D.O., M.C.P. C.E.F., N.C.B.; Writing – Original Draft, C.D.O., M.C.P., C.E.F., N.C.B., E.A.G., and E.A.O.; Writing – Review & Editing, C.D.O., E.A.G., and E.A.O.; Funding Acquisition, E.A.G. and E.A.O.; Resources, E.A.G. and E.A.O; Supervision, E.A.O.

Subjects:

Research Funding:

This work was supported in part by the National Institutes of Health (K12-GM000680 to C.D.O.); W.M. Keck Foundation Medical Research Grant (to E.A.O.).

Crystallographic data were collected at Southeast Regional Collaborative Access Team (SER-CAT) 22-BM beamline at the Advanced Photon Source, Argonne National Laboratory. Use of the Advanced Photon Source was supported by the United States Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract W-31-109-Eng-38.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biochemistry & Molecular Biology
  • Biophysics
  • Cell Biology
  • THERMOPHILIC PROTEINS
  • MOLECULAR-DYNAMICS
  • RESURRECTED PROTEINS
  • SECONDARY STRUCTURE
  • RATIONAL DESIGN
  • NUCLEIC-ACIDS
  • EF-TU
  • STABILITY
  • NETWORKS
  • SOFTWARE

Structural and Dynamics Comparison of Thermostability in Ancient, Modern, and Consensus Elongation Factor Tus

Tools:

Journal Title:

Structure

Volume:

Volume 26, Number 1

Publisher:

, Pages 118-+

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Rationally engineering thermostability in proteins would create enzymes and receptors that function under harsh industrial applications. Several sequence-based approaches can generate thermostable variants of mesophilic proteins. To gain insight into the mechanisms by which proteins become more stable, we use structural and dynamic analyses to compare two popular approaches, ancestral sequence reconstruction (ASR) and the consensus method, used to generate thermostable variants of Elongation Factor Thermo-unstable (EF-Tu). We present crystal structures of ancestral and consensus EF-Tus, accompanied by molecular dynamics simulations aimed at probing the strategies employed to enhance thermostability. All proteins adopt crystal structures similar to extant EF-Tus, revealing no difference in average structure between the methods. Molecular dynamics reveals that ASR-generated sequences retain dynamic properties similar to extant, thermostable EF-Tu from Thermus aquaticus, while consensus EF-Tu dynamics differ from evolution-based sequences. This work highlights the advantage of ASR for engineering thermostability while preserving natural motions in multidomain proteins. Ancestral sequence reconstruction (ASR) and the consensus approach are compared in the generation of thermostable EF-Tu homologs. Using a combination of X-ray crystallography and molecular dynamics simulations, Okafor et al. show that while both methods yield thermostable proteins, ASR, unlike consensus, preserves the natural protein motions in EF-Tu.

Copyright information:

© 2017 Elsevier Ltd

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Creative Commons License

Export to EndNote