Figure A1.

Enzymatic cascade reactions to spectroscopically evaluate L. Hsp90 ATPase kinetics. The ADP produced by ATP hydrolysis during the chaperone cycle reacts with PEP (phosphoenolpyruvate) to form pyruvate in a reaction catalyzed by the enzyme pyruvate kinase (PK), whereby ATP is regenerated. Pyruvate then continues the cascade process and is reduced to lactic acid by action of lactate dehydrogenase (LDH), whereby NADH is oxidized to NAD⁺.