BLV-PR Kinetic parameters determined by using fluorogenic substrates.

(A) Steady-state rate as a function of the CA-NC substrate concentration. BLV-PR (10 nM) was mixed with substrate CA-NC (0.05–20 μM) in reaction buffer, at 37°C, and the steady-state rate of hydrolysis was determined from the linear portion of the fluorescence increase. The black trace represents the best fit of the Michaelis-Menten equation. (B) Steady-state rate as a function of the enzyme concentration. CA-NC substrate (5 μM) was mixed with BLV-PR (0.05–0.2 nM) in the same buffer. The black trace represents the best fit to a linear equation. The data points represent independent steady-state rate measurements for a representative experiment performed in duplicates.