1. The study characterized the effect of a cyclometalated gold(III) compound on the human topoisomerase IB catalytic cycle. 2. Results showed that the gold(III) compound inhibits topoisomerase IB in a dose-dependent manner and the inhibition is irreversible. 3. Mechanism studies found that the gold(III) compound inhibits the topoisomerase IB cleavage step by preventing enzyme-DNA complex formation, but has no effect on the religation step. 4. Electrophoretic mobility shift assays demonstrated that the gold(III) compound directly binds to topoisomerase IB, inhibiting the topoisomerase IB-DNA complex