Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis

Tükenmez, Hasan; Magnussen, Helge Magnus; Kovermann, Michael; Byström, Anders; Wolf-Watz, Magnus

Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis

Dateien

Tükenmez_0-370097.pdf(2.14 MB)

Datum

2016

Autor:innen

Tükenmez, Hasan

Magnussen, Helge Magnus

Kovermann, Michael

Byström, Anders

Wolf-Watz, Magnus

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

PLoS ONE ; 11 (2016), 9. - e0163115. - eISSN 1932-6203

Zusammenfassung

Enzymes have evolved with highly specific values of their catalytic parameters kcat and KM. This poses fundamental biological questions about the selection pressures responsible for evolutionary tuning of these parameters. Here we are address these questions for the enzyme adenylate kinase (Adk) in eukaryotic yeast cells. A plasmid shuffling system was developed to allow quantification of relative fitness (calculated from growth rates) of yeast in response to perturbations of Adk activity introduced through mutations. Biophysical characterization verified that all variants studied were properly folded and that the mutations did not cause any substantial differences to thermal stability. We found that cytosolic Adk is essential for yeast viability in our strain background and that viability could not be restored with a catalytically dead, although properly folded Adk variant. There exist a massive overcapacity of Adk catalytic activity and only 12% of the wild type kcat is required for optimal growth at the stress condition 20°C. In summary, the approach developed here has provided new insights into the evolutionary tuning of kcat for Adk in a eukaryotic organism. The developed methodology may also become useful for uncovering new aspects of active site dynamics and also in enzyme design since a large library of enzyme variants can be screened rapidly by identifying viable colonies.

Fachgebiet (DDC)

540 Chemie

Zitieren

ISO 690

TÜKENMEZ, Hasan, Helge Magnus MAGNUSSEN, Michael KOVERMANN, Anders BYSTRÖM, Magnus WOLF-WATZ, 2016. Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis. In: PLoS ONE. 11(9), e0163115. eISSN 1932-6203. Available under: doi: 10.1371/journal.pone.0163115

BibTex

@article{Tukenmez2016Linka-37467,
  year={2016},
  doi={10.1371/journal.pone.0163115},
  title={Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis},
  number={9},
  volume={11},
  journal={PLoS ONE},
  author={Tükenmez, Hasan and Magnussen, Helge Magnus and Kovermann, Michael and Byström, Anders and Wolf-Watz, Magnus},
  note={Article Number: e0163115}
}

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