Structural basis for ligand binding to an enzyme by a conformational selection pathway

Kovermann, Michael; Grundström, Christin; Sauer-Eriksson, A. Elisabeth; Sauer, Uwe H.; Wolf-Watz, Magnus

Structural basis for ligand binding to an enzyme by a conformational selection pathway

Dateien

Kovermann_2-74zlxgfkgo0a0.pdfGröße: 695.22 KBDownloads: 26

Datum

2017

Autor:innen

Kovermann, Michael

Grundström, Christin

Sauer-Eriksson, A. Elisabeth

Sauer, Uwe H.

Wolf-Watz, Magnus

URI (zitierfähiger Link)

http://nbn-resolving.de/urn:nbn:de:bsz:352-2-74zlxgfkgo0a0

DOI (zitierfähiger Link)

https://doi.org/10.1073/pnas.1700919114

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Chemie
Zukunftskolleg

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Proceedings of the National Academy of Sciences of the United States of America : PNAS. 2017, 114(24), pp. 6298-6303. ISSN 0027-8424. eISSN 1091-6490. Available under: doi: 10.1073/pnas.1700919114

Zusammenfassung

Proteins can bind target molecules through either induced fit or conformational selection pathways. In the conformational selection model, a protein samples a scarcely populated high-energy state that resembles a target-bound conformation. In enzymatic catalysis, such high-energy states have been identified as crucial entities for activity and the dynamic interconversion between ground states and high-energy states can constitute the rate-limiting step for catalytic turnover. The transient nature of these states has precluded direct observation of their properties. Here, we present a molecular description of a high-energy enzyme state in a conformational selection pathway by an experimental strategy centered on NMR spectroscopy, protein engineering, and X-ray crystallography. Through the introduction of a disulfide bond, we succeeded in arresting the enzyme adenylate kinase in a closed high-energy conformation that is on-pathway for catalysis. A 1.9-Å X-ray structure of the arrested enzyme in complex with a transition state analog shows that catalytic sidechains are properly aligned for catalysis. We discovered that the structural sampling of the substrate free enzyme corresponds to the complete amplitude that is associated with formation of the closed and catalytically active state. In addition, we found that the trapped high-energy state displayed improved ligand binding affinity, compared with the wild-type enzyme, demonstrating that substrate binding to the high-energy state is not occluded by steric hindrance. Finally, we show that quenching of fast time scale motions observed upon ligand binding to adenylate kinase is dominated by enzyme-substrate interactions and not by intramolecular interactions resulting from the conformational change.

Fachgebiet (DDC)

540 Chemie

Zitieren

ISO 690

KOVERMANN, Michael, Christin GRUNDSTRÖM, A. Elisabeth SAUER-ERIKSSON, Uwe H. SAUER, Magnus WOLF-WATZ, 2017. Structural basis for ligand binding to an enzyme by a conformational selection pathway. In: Proceedings of the National Academy of Sciences of the United States of America : PNAS. 2017, 114(24), pp. 6298-6303. ISSN 0027-8424. eISSN 1091-6490. Available under: doi: 10.1073/pnas.1700919114

BibTex

@article{Kovermann2017-06-13Struc-39444,
  year={2017},
  doi={10.1073/pnas.1700919114},
  title={Structural basis for ligand binding to an enzyme by a conformational selection pathway},
  number={24},
  volume={114},
  issn={0027-8424},
  journal={Proceedings of the National Academy of Sciences of the United States of America : PNAS},
  pages={6298--6303},
  author={Kovermann, Michael and Grundström, Christin and Sauer-Eriksson, A. Elisabeth and Sauer, Uwe H. and Wolf-Watz, Magnus}
}

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