Substrate binding assay to distinguish if substrate addition is random or ordered. Number of binding sites and K s are determined by a Scatchard Plot . If radioactive sucrose is produced when sucrose plus enzyme are incubated with radioactive fructose, then 1st step must occur. Reaction mechanism is Ping Pong Bi Bi. 4 Radioactive maltose is not produced when radioactive glucose is incubated with the enzyme and maltose.
Reaction mechanism is single displacement.