Fig. 2: Purified CS polymerase complexes are well-folded.
From: Structural basis for human chondroitin sulfate chain polymerization
a Coomassie-stained SDS-PAGE analysis of purified CS polymerase complexes. The expected molecular weight of 3C protease-cleaved proteins is 88 kDa for CHSY1 (aa68-aa802), 86 kDa for CHSY3 (aa157-aa882), 79 kDa for CHPF (aa81-aa775), and 81 kDa for CHPF2 (aa57-aa772). Comparable results were obtained from at least two independent purifications for each complex. b The melting temperature (Tm) of CS polymerase complexes was determined using nano-differential scanning fluorimetry (nanoDSF). The graph corresponds to the first derivative plot of the fluorescence 350 nm/330 nm ratio. Average melting temperatures from technical duplicate measurements are indicated, with corresponding standard deviation in brackets. c–f Mass photometry analysis of purified CS polymerase complexes. The expected molecular masses are 167 kDa for CHSY1–CHPF, 169 kDa for CHSY1–CHPF2, 165 kDa for CHSY3–CHPF, and 167 kDa for CHSY3–CHPF2. Molecular masses from duplicate experiments are shown in the graph; additional measured values are provided in Supplementary Table 2. Source data are provided as a Source Data file.
