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. 1998 Aug 4;95(16):9202–9207. doi: 10.1073/pnas.95.16.9202

Figure 2.

Figure 2

DEPC inactivation of Nuc. (A) Inactivation of Nuc by increasing concentrations of DEPC. Nuc (12 μM) was incubated for 5 min at pH 4.5 with 50 mM DEPC. Hydroxylamine was added to one-half of the reaction at a final concentration of 0.5 M. At the indicated times, the reaction mixtures were diluted 10-fold into assay buffer containing 100 mM pNP-PPh at pH 7.0. Activity was measured for 1 min as described in Experimental Procedures. (B) Time course of inactivation, in presence and absence of tungstate. Nuc (12 μM) was mixed with 50 mM DEPC in the presence or absence (“Control”) of 1 mM tungstate at pH 4.5. Aliquots were removed at the indicated times and diluted 10-fold into assay buffer containing 100 mM pNP-PPh, at pH 7.0, and activity was measured as described.