Studies on the porphobilinogen deaminase-uroporphyrinogen cosynthetase system of cultured soya-bean cells.
1. Porphobilinogenase was isolated and purified from soya-bean callus tissue; its components, porphobilinogen deaminase and uroporphyrinogen isomerase, were separated and purified. 2. The purified porphobilinogenase was resolved into two bands on starch-gel electrophoresis. The molecular weights of porphobilinogenase, deaminase and isomerase fractions were determined by the gel-filtration method. Porphobilinogenase activity was affected by the presence of air; uroporphyrinogens were only formed under anaerobic conditions, although substrate consumption was the same in the absence of oxygen as in its presence. 3. pH-dependence of both porphobilinogenase and deaminase was the same and a sharp optimum at pH 7.2 was obtained. Isomerase was heat-labile, but the presence of ammonium ions or porphobilinogen afforded some protection against inactivation. The action of several compounds added to the system was studied. Cysteine, thioglycollate, ammonium ions and hydroxylamine inhibited porphobilinogenase; certain concentrations of sodium and magnesium salts enhanced activity; some dicarboxylic acids and 2-methoxy-5-nitrotropone inhibited the deaminase. 4. delta-Aminolaevulate and ethionine in the culture media stimulated porphyrin synthesis and increased porphobilinogenase activity, whereas iron deficiency resulted in porphyrin accumulation. 5. The development of chlorophyll and porphobilinogenase on illumination of dark-grown callus was followed. 6. A hypothetical scheme is suggested for the enzymic synthesis of uroporphyrinogens from porphobilinogen.
D000596:Amino Acids / Q000494:pharmacology; D000641:Ammonia / Q000494:pharmacology; D002478:Cells, Cultured / Q000201:enzymology*; D003545:Cysteine / Q000494:pharmacology; D003998:Dicarboxylic Acids / Q000494:pharmacology; D004586:Electrophoresis; D005001:Ethionine / Q000494:pharmacology; D005782:Gels; D006863:Hydrogen-Ion Concentration; D006898:Hydroxylamines / Q000494:pharmacology; D007535:Isomerases; D007982:Levulinic Acids / Q000494:pharmacology; D008190:Lyases; D008274:Magnesium / Q000494:pharmacology; D010100:Oxygen; D010944:Plants / Q000201:enzymology*; D011166:Porphyrins / Q000378:metabolism; D011422:Propionates; D011758:Pyrroles / Q000378:metabolism; D012964:Sodium / Q000494:pharmacology; D013025:Glycine max; D013213:Starch; D013864:Thioglycolates / Q000494:pharmacology
D000596:Amino Acids; D003998:Dicarboxylic Acids; D005782:Gels; D006898:Hydroxylamines; D007982:Levulinic Acids; D011166:Porphyrins; D011422:Propionates; D011758:Pyrroles; D013864:Thioglycolates; D000641:Ammonia; D013213:Starch; D012964:Sodium; D008190:Lyases; D007535:Isomerases; D008274:Magnesium; D003545:Cysteine; D010100:Oxygen; D005001:Ethionine