dc.contributor.authorQuistgaard, Esben M.
dc.contributor.authorWeininger, Ulrich
dc.contributor.authorUral-Blimke, Yonca
dc.contributor.authorModig, Kristofer
dc.contributor.authorNordlund, Pär
dc.contributor.authorAkke, Mikael
dc.contributor.authorLöw, Christian
dc.date.accessioned2016-11-02T06:32:23Z
dc.date.available2016-11-02T06:32:23Z
dc.date.issued2016
dc.identifier.citationQuistgaard, E. M., Weininger, U., Ural-Blimke, Y., Modig, K., Nordlund, P., Akke, M., et al. (2016). Molecular insights into substrate recognition and catalytic mechanism of the chaperone and FKBP peptidyl-prolyl isomerase SlyD. BMC Biology, 14(82), 1-25.en_US
dc.identifier.issn1741-7007en_US
dc.identifier.urihttp://hdl.handle.net/10220/41603
dc.description.abstractBackground: Peptidyl-prolyl isomerases (PPIases) catalyze cis/trans isomerization of peptidyl-prolyl bonds, which is often rate-limiting for protein folding. SlyD is a two-domain enzyme containing both a PPIase FK506-binding protein (FKBP) domain and an insert-in-flap (IF) chaperone domain. To date, the interactions of these domains with unfolded proteins have remained rather obscure, with structural information on binding to the FKBP domain being limited to complexes involving various inhibitor compounds or a chemically modified tetrapeptide. Results: We have characterized the binding of 15-residue-long unmodified peptides to SlyD from Thermus thermophilus (TtSlyD) in terms of binding thermodynamics and enzyme kinetics through the use of isothermal titration calorimetry, nuclear magnetic resonance spectroscopy, and site-directed mutagenesis. We show that the affinities and enzymatic activity of TtSlyD towards these peptides are much higher than for the chemically modified tetrapeptides that are typically used for activity measurements on FKBPs. In addition, we present a series of crystal structures of TtSlyD with the inhibitor FK506 bound to the FKBP domain, and with 15-residue-long peptides bound to either one or both domains, which reveals that substrates bind in a highly adaptable fashion to the IF domain through β-strand augmentation, and can bind to the FKBP domain as both types VIa1 and VIb-like cis-proline β-turns. Our results furthermore provide important clues to the catalytic mechanism and support the notion of inter-domain cross talk. Conclusions: We found that 15-residue-long unmodified peptides can serve as better substrate mimics for the IF and FKBP domains than chemically modified tetrapeptides. We furthermore show how such peptides are recognized by each of these domains in TtSlyD, and propose a novel general model for the catalytic mechanism of FKBPs that involves C-terminal rotation around the peptidyl-prolyl bond mediated by stabilization of the twisted transition state in the hydrophobic binding site.en_US
dc.format.extent25 p.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesBMC Biologyen_US
dc.rights© 2016 The Author(s) (BioMed Central). This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.en_US
dc.subjectPeptidyl-prolyl isomerase (PPIase)en_US
dc.subjectFK506-binding protein (FKBP)en_US
dc.titleMolecular insights into substrate recognition and catalytic mechanism of the chaperone and FKBP peptidyl-prolyl isomerase SlyDen_US
dc.typeJournal Article
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.identifier.doihttp://dx.doi.org/10.1186/s12915-016-0300-3
dc.description.versionPublished versionen_US


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