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. Author manuscript; available in PMC: 2023 Dec 15.
Published in final edited form as: Cell Chem Biol. 2022 Nov 23;29(12):1680–1693.e9. doi: 10.1016/j.chembiol.2022.11.003

Figure 4. Surface region around C66 is an allosteric binding site of RSL3 on GPX4.

Figure 4.

A, Intact protein MALDI MS of GPX4U46C or GPX4U46C-C66S preincubated with DMSO or RSL3. B, Enzymatic activity of GPX4U46C and GPX4U46C_C66S preincubated with DMSO, RSL3 (20 μM), or ML162 (20 μM) were acquired (n=3). Data are represented as mean ± s.d. One-way ANOVA followed by Dunnett’s multiple comparisons test was performed: pns>0.05 and p** < 0.01. C, Backbone assignments of 1H, 15N-HSQC-NMR spectrum for 15N-GPX4U46C. Horizontal red lines point out the peaks corresponding to Asparagine and Glutamine side chain NH2. D, Overlap of 1H, 15N-HSQC-NMR spectrum of 50 μM 15N-GPX4U46C alone and its mixture with 100 μM RSL3 (both in 5.0 % d6-DMSO), with zoom-in panels on selected peaks of prominent changes.

See also Figure S4.