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. Author manuscript; available in PMC: 2021 May 1.
Published in final edited form as: J Steroid Biochem Mol Biol. 2019 Dec 20;199:105567. doi: 10.1016/j.jsbmb.2019.105567

Figure 6. Total ion count chromatographs of rDesAB reaction products.

Figure 6.

Reaction was initiated with 2 μM of pure rDesAB enzyme to 50 mM MOPS buffer (pH 7.0), 100 μM of substrate (blue) in the presence of 1 μM MnCl2 and 10 μM TPP at room temperature. After 2 minutes, while the reactions were stopped with the addition of 1/10 volume of 1 M HCl and vortexing to drop the pH below 2. A final concentration of 50 uM of internal standard, 20β-dihydrocortisol (green), was spiked into the reaction. Products formed were identical with respect to retention time and loss of 60 amu consistent with side-chain cleavage.