Purification and properties of ferrochelatase from Chironomidae larvae.
Ferrochelatase with an Mr of 42,700 Da and a pI of 7.35 has been purified to homogeneity from chironomidae larvae. The activity of the enzyme reached maximum at pH 7.8 and decreased with the increase of pH. The enzyme activity varied with temperature and showed maximum activity around 37 degrees C. The purified enzyme was active towards protoporphyrin but inactive towards other porphyrins. The specific enzyme activity of ferrochelatase from chironomidae is about 10-fold higher than that of the rat. Electrophoresis of the purified fractions shows that the enzyme contains only one single polypeptide. The soluble ferrochelatase contained one mole of iron in each mole of the enzyme. The N-terminal sequence analysis of the enzyme shows a high percentage of conserved regions of the enzyme among other species. The enzyme properties are similar to those of the mammalian ferrochelatases except with slightly higher specific activity. Chironomidae ferrochelatase appeared to be more heat resistant and less susceptible than its mammalian equivalent to inhibition by lead.
Leung
Y K
YK
Department of Biochemistry, The Chinese University of Hong Kong, Shatin.
Ho
J W
JW
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000645:Ammonium Sulfate / Q000494:pharmacology; D000818:Animals; D002683:Chironomidae / Q000196:embryology* / Q000201:enzymology*; D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D017124:Conserved Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005294:Ferrochelatase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D006418:Heme / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000201:enzymology; D007501:Iron / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011166:Porphyrins / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011524:Protoporphyrins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D000596:Amino Acids; D010455:Peptides; D011166:Porphyrins; D011524:Protoporphyrins; D006418:Heme; D007501:Iron; D005294:Ferrochelatase; D000645:Ammonium Sulfate