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. Author manuscript; available in PMC: 2022 May 10.
Published in final edited form as: Nat Struct Mol Biol. 2021 Nov 10;28(11):923–935. doi: 10.1038/s41594-021-00677-4

Figure 2. The RGG3 and SYGQ LC remain disordered in a two-component condensed phase.

Figure 2.

A) A two-component phase consisting of 2:1 13C-labeled SYGQ LC and 15N-labeled RGG3 was made by inducing phase separation of the two components and centrifuging the sample to create a liquid condensed phase within an NMR tube. B) 1H-15N HSQC of the two-component condensed phase overlaid with a spectrum of dispersed phase (100 μM) RGG3. Asterisks denote resonances arising from natural isotopic abundance glutamine side chains within the SYGQ LC. C) Chemical shift perturbations of RGG3 in the condensed phase relative to the dispersed phase. Certain resonances are not included due to lack of resolution or signal broadening. D) 15N spin relaxation values comparing local reorientational motions of RGG3 in the condensed (purple) and dispersed (orange) phases. Data are plotted as mean ± propagated best-fit parameter confidence interval equal to one s.d. in one out of two independent experiments. E) 1H-13C HSQC of the two-component condensed phase overlaid with a condensed phase of the SYGQ LC alone from prior work19. Asterisks correspond to resonances from the initiator methionine present only within the protein construct used in this study.