Danio rerio Oocytes for Eukaryotic In-Cell NMR.

Joseph F Thole, Tanner C Fadero, Jeffrey P Bonin, Samantha S Stadmiller, Jonathan A Giudice, Gary J Pielak

Biochemistry 2021 Feb 16

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Understanding how the crowded and complex cellular milieu affects protein stability and dynamics has only recently become possible by using techniques such as in-cell nuclear magnetic resonance. However, the combination of stabilizing and destabilizing interactions makes simple predictions difficult. Here we show the potential of Danio rerio oocytes as an in-cell nuclear magnetic resonance model that can be widely used to measure protein stability and dynamics. We demonstrate that in eukaryotic oocytes, which are 3-6-fold less crowded than other cell types, attractive chemical interactions still dominate effects on protein stability and slow tumbling times, compared to the effects of dilute buffer.

Citation

Joseph F Thole, Tanner C Fadero, Jeffrey P Bonin, Samantha S Stadmiller, Jonathan A Giudice, Gary J Pielak. Danio rerio Oocytes for Eukaryotic In-Cell NMR. Biochemistry. 2021 Feb 16;60(6):451-459