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. Author manuscript; available in PMC: 2012 Nov 7.
Published in final edited form as: J Biol Chem. 2004 Apr 21;279(28):29493–29500. doi: 10.1074/jbc.M403187200

Figure 4. Determination of stoichiometry of alkylation by quinacrine mustard.

Figure 4

A sample of NADPH-reduced TryR (1 μM) was inactivated by 10 μM QM. Aliquots were removed at intervals, quenched with 2-mercaptoethanol and residual activities were determined as described in Figure 2. Residual activity is fitted to the equation for a single exponential decay (open circles). The remainder of each quenched sample was denatured with trichloroacetic acid to remove bound FAD, washed and re-dissolved by proteolytic digestion. Residual covalently bound QM was determined spectrophotometrically as described in Experimental Procedures (closed circles).