Journal: Journal of Lipid Research

Article Title: Structure of mouse cytosolic sulfotransferase SULT2A8 provides insight into sulfonation of 7α-hydroxyl bile acids

doi: 10.1016/j.jlr.2021.100074

Figure Lengend Snippet: SULT2A8 stability in the presence of cofactor and substrates. A: Trypsin limited proteolysis study. Purified SULT2A8 was coincubated with or without PAP and Na CDC as indicated in the trypsin digestion reactions. Digested proteins were electrophoresed by SDS-PAGE and detected by Coomassie blue staining. B: SULT2A8 WT and its mutants with substitution in the active site (upper panel) or loop 3 (lower panel) were coincubated with PAP, native/non-native substrates as indicated. The reaction was assayed using JBS Thermofluor Dye system. Fluorescent at 568 nm was detected with excitation wavelength of 483 nm under heating in a ramp rate of 1°C/min. T m was calculated by solving the nonlinear fit through a Boltzmann sigmoidal equation and presented in means and standard deviation (n = 3). Differences of each ligand-treated protein were compared with its corresponding apo form using unpaired Student’s t -test, and significance was indicated in the text.

Article Snippet: This was followed by mixing 20 μl of prepared protein with 5 μl of 500-fold diluted JBS Thermofluor Dye (Jena Bioscience) in the same buffer.

Techniques: Purification, SDS Page, Staining, Standard Deviation