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Alanine to serine substitutions drive thermal adaptation in a psychrophilic diatom cytochrome c6

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Abstract

In this study, we investigate the thermodynamic mechanisms by which electron transfer proteins adapt to environmental temperature by directly comparing the redox properties and folding stability of a psychrophilic cytochrome c and a mesophilic homolog. Our model system consists of two cytochrome c6 proteins from diatoms: one adapted specifically to polar environments, the other adapted generally to surface ocean environments. Direct electrochemistry shows that the midpoint potential for the mesophilic homolog is slightly higher at all temperatures measured. Cytochrome c6 from the psychrophilic diatom unfolds with a melting temperature 10.4 °C lower than the homologous mesophilic cytochrome c6. Changes in free energy upon unfolding are identical, within error, for the psychrophilic and mesophilic protein; however, the chemical unfolding transition of the psychrophilic cytochrome c6 is more cooperative than for the mesophilic cytochrome c6. Substituting alanine residues found in the mesophile with serine found in corresponding positions of the psychrophile demonstrates that burial of the polar serine both decreases the thermal stability and decreases the midpoint potential. The mutagenesis data, combined with differences in the m-value of chemical denaturation, suggest that differences in solvent accessibility of the hydrophobic core underlie the adaptation of cytochrome c6 to differing environmental temperature.

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Abbreviations

cyt c :

Cytochrome c

Fc:

Fragilariopsis cylindrus

Tp:

Thalassiosira pseudonana

PGE:

Pyrolytic graphite edge

Gdn:

Guanidine HCl

E mid :

Midpoint potential

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Acknowledgements

The authors thank the M. J. Murdock Charitable Trust for funding through the Murdock College Research Program for Natural Sciences. We thank the Seattle University College of Science and Engineering for summer support. Seattle University students enrolled in CHEM 1910 collected key preliminary cyclic voltammetry data for this study.

Funding

This study was funded by the M. J. Murdock Charitable Trust Murdock College Research Program for Natural Sciences (2015302:MNL).

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  1. Department of Chemistry, Seattle University, 901 12th Ave., Seattle, WA, 98122, USA

    Miranda Wilson, Logan Tillery, Emily Tabaie, Galen Beery, Jordyn Preusker, Inaara Bhola & Katherine Frato

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  1. Miranda Wilson
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Wilson, M., Tillery, L., Tabaie, E. et al. Alanine to serine substitutions drive thermal adaptation in a psychrophilic diatom cytochrome c6. J Biol Inorg Chem 25, 489–500 (2020). https://doi.org/10.1007/s00775-020-01777-0

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