Abstract

Recombinant wild-type human liver arginase (EC 3.5.3.1) expressed in Escherichia coli was markedly resistant to inhibition by ethylene diamine tetraacetic acid (EDTA). In contrast, half and fully activated species of the H101N variant were totally inactive in the presence of similar to 1 mM EDTA. Dilution of inhibited species in metal-free buffer lead to a time dependent recovery of activity, even when measured in the absence of added Mn2+. The inhibition was mixed type, with predominance of a competitive component (K-ii = 0.31 mM; K-is = 0.022 mM). The structurally related N, N, N, N'-tetramethylethylenediamine was not inhibitory, indicating the importance of the carboxyl groups in EDTA inhibition. We conclude that EDTA inhibition of H101N arginase is not due to interaction with a weakly bound Mn2+ or chelation of essential metal ions. (C) 2004 Elsevier Inc. All rights reserved.