X-ray structure and characterization of carbamate kinase from the human parasite Giardia lamblia

Carbamate kinase catalyzes the reversible conversion of carbamoyl phosphate and ADP to ATP and ammonium carbamate, which is hydrolyzed to ammonia and carbonate. The three-dimensional structure of carbamate kinase from the human parasite Giardia lamblia (glCK) has been determined at 3 Å resolution. The crystals belonged to the monoclinic space group P2₁, with unit-cell parameters a = 69.77, b = 85.41, c = 102.1 Å, β = 106.8°. The structure was refined to a final R factor of 0.227. The essentiality of glCK together with its absence in humans makes the enzyme an attractive candidate for anti-Giardia drug development. Steady-state kinetic rate constants have been determined. The k_cat for ATP formation is 319 ± 9 s⁻¹. The K_m values for carbamoyl phosphate and ADP are 85 ± 6 and 70 ± 5 µM, respectively. The structure suggests that three invariant lysine residues (Lys131, Lys216 and Lys278) may be involved in the binding of substrates and phosphoryl transfer. The structure of glCK reveals that a glycerol molecule binds in the likely carbamoyl phosphate-binding site.