How do you analyze multi-substrate enzyme kinetics?

Multi-substrate reactions require expanded kinetic analysis. Mechanisms include: ordered (substrates bind in specific sequence), random (either substrate can bind first), and ping-pong (one product released before second substrate binds). Determining mechanism: initial velocity patterns at varied substrate concentrations, product inhibition patterns, and isotope exchange studies. Analysis: Cleland nomenclature describes kinetic mechanisms systematically. For bisubstrate reactions: v = Vmax[A][B]/(KiaKmB + KmA[B] + KmB[A] + [A][B]) for ordered mechanism. Product inhibition distinguishes ordered (competitive and noncompetitive patterns depending on order) from random (all noncompetitive) mechanisms. Dead-end inhibitor studies provide additional mechanistic information. Understanding mechanism is essential for interpreting kinetic data and designing inhibitors.