An optimized strategy to measure protein stability highlights differences between cold and hot unfolded states

Nature Communications
Caterina AlfanoPiero Andrea Temussi

Abstract

Macromolecular crowding ought to stabilize folded forms of proteins, through an excluded volume effect. This explanation has been questioned and observed effects attributed to weak interactions with other cell components. Here we show conclusively that protein stability is affected by volume exclusion and that the effect is more pronounced when the crowder's size is closer to that of the protein under study. Accurate evaluation of the volume exclusion effect is made possible by the choice of yeast frataxin, a protein that undergoes cold denaturation above zero degrees, because the unfolded form at low temperature is more expanded than the corresponding one at high temperature. To achieve optimum sensitivity to changes in stability we introduce an empirical parameter derived from the stability curve. The large effect of PEG 20 on cold denaturation can be explained by a change in water activity, according to Privalov's interpretation of cold denaturation.

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Citations

Dec 12, 2018·Bioinformatics·Mattia MiottoEdoardo Milanetti
Sep 22, 2020·Journal of Biomolecular Structure & Dynamics·K Tejaswi Naidu, N Prakash Prabhu
Feb 10, 2019·Applied Biochemistry and Biotechnology·Sumra ShahidAsimul Islam
Jul 8, 2020·International Journal of Molecular Sciences·Xiaomin YangSandrine Lacombe
May 28, 2019·Frontiers in Molecular Biosciences·Oliver BrylskiJonathan W Mueller
Mar 16, 2021·Communications Chemistry·Rita PuglisiPiero A Temussi
Apr 4, 2020·Current Opinion in Chemical Biology·Vangelis AgouridasOleg Melnyk
Nov 5, 2020·The Journal of Physical Chemistry. B·K Tejaswi NaiduN Prakash Prabhu
Jun 23, 2020·Biochemistry·Caitlin M Davis, Martin Gruebele
Dec 20, 2018·Journal of the American Chemical Society·Annalisa PastorePiero Andrea Temussi
Sep 23, 2021·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Anastasia S PolitouPiero Andrea Temussi

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Methods Mentioned

BETA
circular dichroism
nuclear magnetic resonance
NMR
electrophoresis

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