Studies on uroporphyrinogen biosynthesis in pig liver.
Porphobilinogen-deaminase (PBG-D) and PBG-D-isomerase complex (PBG-D-I) from pig liver were isolated and partially purified. Uroporphyrinogen I and III formation was found to be linear with time and protein concentration. Optimal pH was about 7.4 and 7.6-7.8 for PBG-D and PBG-D-I complex, respectively. Some properties of the isolated enzymes were studied. Molecular mass determination gave a value of 40,000 Da for PBG-D and 50,000 Da for the complex. Both enzymes exhibited classical Michaelis-Menten kinetics. Km and Vmax parameters were estimated. The effect of several divalent cations, ammonia and thiol reagents was also investigated. The differential action of some of these chemicals on PBG-D and PBG-D-I system would suggest that PBG-D and isomerase may not be only physically adjacent but actually associated.
Fumagalli
S A
SA
Centro de Investigaciones sobre Porfirinas y Porfirias-CIPYP (CONICET, F.C.E. y N., UBA)-Ciudad Universitaria, Buenos Aires, Argentina.
Kotler
M L
ML
Rossetti
M V
MV
Batlle
A M
AM
D000641:Ammonia / Q000494:pharmacology; D000818:Animals; D002412:Cations; D002850:Chromatography, Gel; D011163:Hydroxymethylbilane Synthase / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013552:Swine; D014576:Uroporphyrinogen III Synthetase / Q000302:isolation & purification / Q000378:metabolism*; D014577:Uroporphyrinogens / Q000096:biosynthesis*
D002412:Cations; D009097:Multienzyme Complexes; D013439:Sulfhydryl Reagents; D014577:Uroporphyrinogens; D000641:Ammonia; D011163:Hydroxymethylbilane Synthase; D014576:Uroporphyrinogen III Synthetase