Dissociation and reassociation of immobilized porphobilinogen synthase: use of immobilized subunits for enzyme isolation.
The dissociation and association of an immobilized preparation of the octameric enzyme porphobilinogen synthase [5-aminolevulinate hydro-lyase (adding 5-aminolevulinate and cyclizing), EC 4.2.1.24] is described. On treatment of the immobilized preparation with 4 M urea, four subunits per octamer are removed which can be reassociated into a soluble octameric enzyme. The tetrameric bound residual protein can also be reassembled into an octameric structure, with the same initial enzyme activity, by exposing the residual bound protein to a soluble pure enzyme preparation or to a crude liver extract in the presence of urea. The dissociation of the reconstituted bound enzyme releases subunits that again can be reassembled into a soluble octameric pure protein even when the crude liver preparation is used as the donor of the subunits. Thus, a pure enzyme can be isolated in a reassociation-dissociation cycle. The use of immobilized preparations of oligomeric proteins is considered for intra- and interspecies hybridization studies and for the ready preparation of purified enzyme preparations from different species and is suggested as a model for study of the formation of an oligomeric enzyme in the presence of other polypeptides.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D004800:Enzymes, Immobilized / Q000302:isolation & purification / Q000378:metabolism; D006836:Hydro-Lyases / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D000623:Porphobilinogen Synthase / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding
D004800:Enzymes, Immobilized; D046911:Macromolecular Substances; D006836:Hydro-Lyases; D000623:Porphobilinogen Synthase
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