Fig. 4: PINCHs induce target polymerization in vitro.
From: A pharmacological modality to sequester homomeric proteins
a, DLS experiments following recombinant Keap1 BTB (50 μM; 100 μM binding sites) and EL133 (50 μM; 100 μM binding moieties) over time at room temperature. eq., equivalents. b, WB of EL133-treated (20 h) OCI-AML2 cells with or without treatment with excess BDX (5 μM). Representative results from two independent experiments are shown. c, Additional Keap1 PINCH WB results in OCI-AML2 cells (20 h) (representative result from two independent experiments). d, DLS experiments after treatment with 50 μM recombinant Keap1 BTB with these PINCHs at 50 μM, over time at room temperature. e, Chemical structure of LDHA-targeting PINCH OS-47B. f, DLS experiments following recombinant LDHA incubated with OS-47B over time at room temperature. The reported concentrations indicate binding sites and binding moieties. Each LDHA tetramer comprises four binding sites and each PINCH comprises two binding moieties. We report the concentrations of monomers (for example, 5 μM LDHA and 20 μM PINCH are indicated as 20 μM:40 μM binding sites:binding moieties). g, Chemical structures of NQO2-targeting PINCHs. h,i, DLS experiments following recombinant NQO2 incubated with EL245 (h) or EL244 (i) over time at room temperature. NQO2 is dimeric (two binding sites) and the concentrations are reported in the same way for LDHA (binding sites:binding moieties).
