K. J. Lee, D. H. Kim/Bioorg. Med. Chem. 6 (1998) 1613±1622
1621
buer for 10 min was added an inactivator (®nal con-
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was incubated at room temperature. At speci®ed inter-
vals, 50 mL samples of the incubation mixture were
removed and added to a 950 mL assay mixture and the
activity was monitored at 320 nm (Figure 3).
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Determination of the partition ratios
A series of solutions (500 mL each) containing 14.5 mM of
CPA and various molar equivalents of inactivators were
prepared to give [I]o/[E]o ratios of 0 to 1200 in 0.05 M
Tris/0.5 M NaCl, pH 7.5 buer, supplemented with
15% dimethyl sulfoxide and were stirred gently at 4 ꢀC
for 16 h. The incubation mixtures were then dialyzed in
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Acknowledgements
The authors wish to express their sincere thanks to the
Pohang University of Science and Technology and the
Korea Science and Engineering Foundation for the
®nancial support of this work.
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