title: The role of zinc with special reference to the essential thiol groups in delta-aminolevulinic acid dehydratase of bovine liver.
abstract: delta-Aminolevulinic acid dehydratase (5-aminolevulinic acid hydro-lyase (adding 5-aminolevulinic acid and cyclizing), EC 4.2.1.24 purified from bovine liver in the presence of both SH-reducing reagent and zinc during the purification contained one zinc atom and eight SH groups/subunit. This preparation showed the full enzymatic activity even in the absence of thiol activator. It was found that two cysteine residues, one zinc atom and two histidine residues were involved in the active site. The enzyme was fullly active as long as two SH groups in the active site remained in the reduced form even in the absence of zinc. However, the enzymatic activity was completely lost, with a concomitant loss of bound zinc, upon oxidation of the SH groups to a disulfide bond, modification of SH groups with chemical reagents, or mercaptide formation by heavy metals. Thus, it is apparent that the activity depends on the essential SH groups. The zinc is not absolutely essential for the activity but may be required to prevent the essential SH groups from autooxidation by coordination. Binding experiments indicated that there was one binding site of zinc/subunit. Photooxidation of histidine residues diminished both enzymatic activity and bound zinc, suggesting that the histidine residues not only constituted the active site but also served as a possible ligand to zinc.
mesh_terms: D000818:Animals; D001051:Apoenzymes / Q000378:metabolism; D001665:Binding Sites; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D006639:Histidine; D008099:Liver / Q000201:enzymology*; D010100:Oxygen / Q000494:pharmacology; D000623:Porphobilinogen Synthase / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
chemical_list: D001051:Apoenzymes; D013438:Sulfhydryl Compounds; D006639:Histidine; D000623:Porphobilinogen Synthase; D015032:Zinc; D010100:Oxygen