Crystal cryocooling distorts conformational heterogeneity in a model michaelis complex of DHFR

Daniel A. Keedy, Henry Van Den Bedem, David A. Sivak, Gregory A. Petsko, Dagmar Ringe, Mark A Wilson, James S. Fraser

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Summary Most macromolecular X-ray structures are determined from cryocooled crystals, but it is unclear whether cryocooling distorts functionally relevant flexibility. Here we compare independently acquired pairs of high-resolution data sets of a model Michaelis complex of dihydrofolate reductase (DHFR), collected by separate groups at both room and cryogenic temperatures. These data sets allow us to isolate the differences between experimental procedures and between temperatures. Our analyses of multiconformer models and time-averaged ensembles suggest that cryocooling suppresses and otherwise modifies side-chain and main-chain conformational heterogeneity, quenching dynamic contact networks. Despite some idiosyncratic differences, most changes from room temperature to cryogenic temperature are conserved and likely reflect temperature-dependent solvent remodeling. Both cryogenic data sets point to additional conformations not evident in the corresponding room temperature data sets, suggesting that cryocooling does not merely trap preexisting conformational heterogeneity. Our results demonstrate that crystal cryocooling consistently distorts the energy landscape of DHFR, a paragon for understanding functional protein dynamics.

Original languageEnglish (US)
Pages (from-to)899-910
Number of pages12
JournalStructure
Volume22
Issue number6
DOIs
StatePublished - Jun 10 2014

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Tetrahydrofolate Dehydrogenase
Temperature
X-Rays
Datasets
Proteins

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Keedy, D. A., Van Den Bedem, H., Sivak, D. A., Petsko, G. A., Ringe, D., Wilson, M. A., & Fraser, J. S. (2014). Crystal cryocooling distorts conformational heterogeneity in a model michaelis complex of DHFR. Structure, 22(6), 899-910. https://doi.org/10.1016/j.str.2014.04.016

Crystal cryocooling distorts conformational heterogeneity in a model michaelis complex of DHFR. / Keedy, Daniel A.; Van Den Bedem, Henry; Sivak, David A.; Petsko, Gregory A.; Ringe, Dagmar; Wilson, Mark A; Fraser, James S.

In: Structure, Vol. 22, No. 6, 10.06.2014, p. 899-910.

Research output: Contribution to journalArticle

Keedy, DA, Van Den Bedem, H, Sivak, DA, Petsko, GA, Ringe, D, Wilson, MA & Fraser, JS 2014, 'Crystal cryocooling distorts conformational heterogeneity in a model michaelis complex of DHFR', Structure, vol. 22, no. 6, pp. 899-910. https://doi.org/10.1016/j.str.2014.04.016
Keedy, Daniel A. ; Van Den Bedem, Henry ; Sivak, David A. ; Petsko, Gregory A. ; Ringe, Dagmar ; Wilson, Mark A ; Fraser, James S. / Crystal cryocooling distorts conformational heterogeneity in a model michaelis complex of DHFR. In: Structure. 2014 ; Vol. 22, No. 6. pp. 899-910.
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