Zinc and other divalent cations inhibit purified leukotriene A4 hydrolase and leukotriene B4 biosynthesis in human polymorphonuclear leukocytes.
Leukotriene A4 hydrolase is a bifunctional metalloenzyme that contains 1 mol of zinc per mole of protein. The primary function of the metal is catalytic and zinc is thus necessary for both its peptidase and its epoxide hydrolase activity. However, at concentrations of zinc exceeding a 1:1 molar ratio (metal:enzyme), we found that zinc acted as an inhibitor with IC50 values of 10 microM for the epoxide hydrolase activity, i.e., the conversion of leukotriene A4 to leukotriene B4, and 0.1 microM for the peptidase activity. The inhibition of both enzyme activities could be reversed by treating the enzyme with chelating agents such as EDTA or dipicolinic acid. Several divalent cations, other than zinc, were also found to inhibit leukotriene A4 hydrolase although with different specificity and potency for the two enzyme activities. Thus, CdSO4 and HgCl2 were effective inhibitors (IC50 approximately 10 microM) of the epoxide hydrolase activity, whereas CoCl2 or MnCl2 were not inhibitory even at concentrations of 1 mM. On the other hand, the peptidase activity was inhibited by CdSO4, NiSO4, HgCl2, MnCl2, CoCl2, and PbNO3, listed in decreasing order of potencies (IC50 0.5-10 microM). In addition, zinc in micromolar concentrations inhibited leukotriene B4 formation in intact human polymorphonuclear leukocytes stimulated by the calcium ionophore A23187 and cell homogenates incubated with arachidonic acid. However, this effect was not related to inhibition of leukotriene A4 hydrolase but rather to a direct or indirect inhibitory effect on the enzyme 5-lipoxygenase in isolated leukocytes. In these cells, 15-lipoxygenase activity was also inhibited by zinc (IC50 5 microM), whereas leukotriene C4 synthase activity in human platelets and rat basophilic leukemia cells was significantly affected only at concentrations > or = 1 mM.
Wetterholm
A
A
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Macchia
L
L
Haeggström
J Z
JZ
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology / Q000378:metabolism; D002104:Cadmium / Q000494:pharmacology; D019187:Cadmium Compounds; D002413:Cations, Divalent / Q000494:pharmacology*; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D010450:Endopeptidases / Q000097:blood* / Q000378:metabolism; D004851:Epoxide Hydrolases / Q000037:antagonists & inhibitors / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D007700:Kinetics; D015471:Leukemia, Basophilic, Acute; D007975:Leukotriene B4 / Q000096:biosynthesis* / Q000097:blood; D008627:Mercuric Chloride / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D051381:Rats; D013431:Sulfates / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology*; D017967:Zinc Compounds / Q000494:pharmacology; D019287:Zinc Sulfate
D019187:Cadmium Compounds; D002413:Cations, Divalent; D002614:Chelating Agents; D013431:Sulfates; D017967:Zinc Compounds; D002104:Cadmium; D007975:Leukotriene B4; D008627:Mercuric Chloride; D019287:Zinc Sulfate; C037123:cadmium sulfate; D004851:Epoxide Hydrolases; D010450:Endopeptidases; D015032:Zinc; C043134:leukotriene A4 hydrolase