Order of uroporphyrinogen III decarboxylation on incubation of porphobilinogen and uroporphyrinogen III with erythrocyte uroporphyrinogen decarboxylase.
The isomeric compositions of the heptacarboxylic, hexacarboxylic and pentacarboxylic porphyrinogens formed by incubation of porphobilinogen with human red-cell haemolysates have been analysed and compared with those derived from incubation with chemically prepared uroporphyrinogen III as substrate. The results indicated that when supplied with an excess (3.7 microM) of exogenous uroporphyrinogen III, uroporphyrinogen decarboxylase utilized the substrate at random and a mixture of isomers was produced; whereas with uroporphyrinogen III generated enzymically from porphobilinogen as substrate a clockwise decarboxylation sequence was observed, resulting in the formation of intermediates mainly with the ring-D, rings-AD and rings-ABD acetate groups decarboxylated. Using [14C]uroporphyrinogen III as substrate at low concentrations (0.01-0.5 microM) also led to preferential decarboxylation of the ring-D acetate group. It was concluded that the order of uroporphyrinogen III decarboxylation is substrate-concentration-dependent, and under normal physiological conditions enzymic decarboxylation is most probably orderly and clockwise, starting at the ring-D acetate group.
Luo
J
J
MRC Toxicology Unit, Medical Research Council Laboratories, Carshalton, Surrey, U.K.
Lim
C K
CK
D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D011162:Porphobilinogen / Q000097:blood*; D011166:Porphyrins / Q000302:isolation & purification / Q000378:metabolism; D014575:Uroporphyrinogen Decarboxylase / Q000097:blood*; D014577:Uroporphyrinogens / Q000097:blood*
D011166:Porphyrins; D014577:Uroporphyrinogens; D011162:Porphobilinogen; D014575:Uroporphyrinogen Decarboxylase