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. 2022 Feb 17;298(4):101739. doi: 10.1016/j.jbc.2022.101739

Figure 3.

Figure 3

Assessing the effects of common buffer conditions on Mproinitial rate. All tests are done in 100 μl buffer containing 100 μM VKLQ–AMC substrate and 200 nM enzyme. A, pH optimization in 20 mM NaPO4 (pH 6.0–8.0) with 150 mM NaCl. B, preference of Mpro for Hepes, Tris, Bis–Tris, or NaPO4, in 20 mM buffering agent, pH 7.0, with 150 mM NaCl. C, preference of Mpro for Tris, Bis–Tris, and NaPO4 in 20 mM buffering agent, pH 7.0. D, effect of 0 to 300 mM NaCl in 20 mM NaPO4 (pH 7.0). E, effect of 0 to 30% v/v glycerol in 20 mM NaPO4 (pH 7.0) with 150 mM NaCl. F, effect of 1 to 10% v/v DMSO in 20 mM NaPO4 (pH 7.0) with 150 mM NaCl. Each measurement is reported as the mean with error bars showing ±1 standard deviation, n = 3. AMC, 7-amino-4-methylcoumarin; DMSO, dimethyl sulfoxide; Mpro, main protease.