Studies on the active centre of rat liver porphyrinogen carboxylase in vivo effect of hexachlorobenzene.
1. Porphyrinogen carboxylase from the liver of normal and hexachlorobenzene porphyric rats was subjected to chemical modification using photo-oxidation with methylene blue, diethylpyrocarbonate, butane-2,3-dione, and phenylglyoxal. 2. All of these chemicals inactivated the enzyme from both sources. 3. Reversion of the diethylpyrocarbonate reaction with hydroxylamine as well as protection of the enzymes with uroporphyrinogen III indicated that histidine is involved at least in the first decarboxylation active site of the porphyrinogen carboxylyase, and perhaps in one or more sites where the removal of the other carboxyl groups take place. 4. Arginine seems not to be at the active site of the enzyme but at its environment since two diketones alter the enzyme activity, however the substrate did not protect the enzyme from the butane-2,3-dione modification. 5. Comparative studies between the enzyme from normal and porphyric animals suggest that the low enzyme activity from intoxicated animals could be due to alterations of its active centre environment produced by hexachlorobenzene treatment. This treatment seems to partially protect the active site of the porphyrinogen carboxylase from the modification reactions.
Billi de Catabbi
S
S
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina.
Ríos de Molina
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MC
San Martín de Viale
L C
LC
D000818:Animals; D001120:Arginine; D001665:Binding Sites; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D004852:Epoxy Compounds / Q000494:pharmacology; D005260:Female; D006581:Hexachlorobenzene / Q000494:pharmacology*; D006639:Histidine; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D008099:Liver / Q000201:enzymology*; D008751:Methylene Blue; D010777:Photochemistry; D011164:Porphyrias / Q000139:chemically induced / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry; D014577:Uroporphyrinogens / Q000494:pharmacology
D004852:Epoxy Compounds; D006898:Hydroxylamines; D014577:Uroporphyrinogens; D019811:Hydroxylamine; D006639:Histidine; D006581:Hexachlorobenzene; C007366:diepoxybutane; D001120:Arginine; D002262:Carboxy-Lyases; C022430:porphyrinogen carboxy-lyase; D004047:Diethyl Pyrocarbonate; D008751:Methylene Blue