LEUKOTRIENE-A4 HYDROLASE - AN ANION ACTIVATED PEPTIDASE

The peptidase activity of leukotriene A4 hydrolase purified from human leukocytes has been characterized, utilizing synthetic amides as substrates. The enzyme was stimulated by several monovalent anions. Thiocyanate ions were most effective followed by chloride and bromide ions. In phosphate buffer alone the peptidase activity towards alanine-4-nitroanilide was barely detectable and addition of 100 mM NaCl increased the specific activity more than 20-fold. Increasing the concentration of NaCl (or NaSCN) did not significantly affect the apparent K(m) for the substrate alanine-4-nitroanilide, but resulted in a dose dependent increase of V(max). The stimulatory effect of these anions on the reaction velocities appeared to obey saturation kinetics and thus indicated the presence of an anion binding site. Apparent affinity constants for chloride and thiocyanate ions were calculated to 100 and 50 mM, respectively. In contrast to the effect on the peptidase activity, no chloride-stimulation could be detected of the epoxide hydrolase activity of this enzyme, i.e., the conversion of leukotriene A4 into leukotriene B4. In conclusion, the results indicate that under physiological conditions, chloride ions may selectively stimulate the peptidase activity of LTA4 hydrolase. Also, the differences in chloride concentrations between cellular compartments suggest that a possible proteolytic function of the enzyme may be limited to the extracellular space.