An effective chromatographic separation of chicken red blood cell coproporphyrinogen oxidase and uroporphyrinogen decarboxylase, two enzymes in heme biosynthesis.
Of the heme biosynthetic pathway enzymes, coproporphyrinogen oxidase is one of the least understood. Substrate recognition studies [Prepr. Biochem. Biotech.1997, 27, 47, J. Org. Chem.1999, 64, 464] have been done using chicken blood hemolysates (CBH) as the source of this enzyme. However, the enzyme uroporphyrinogen decarboxylase is also present in these preparations and separation of these two enzymes from CBH had not yet been achieved. Thus, a substrate ligand column was developed by covalently linking coproporphyrin-III to a sepharose resin following a similar procedure previously used for the purification of uroporphyrinogen decarboxylase [Int. J. Biochem.1992, 24, 105]. The ligand-resin chromatography step rapidly separates coproporphyrinogen oxidase from uroporphyrinogen decarboxylase as well as the majority of the hemoglobin.
Jones
Marjorie A
MA
Department of Chemistry, Illinois State University, Normal, IL 61790-4160, USA. mjones@xenon.che.ilstu.edu
Taneja
Munish
M
Xu
Yan
Y
Chung
Wen
W
Stob
Christian M
CM
Lash
Timothy D
TD
D000818:Animals; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003304:Coproporphyrinogen Oxidase / Q000737:chemistry* / Q000302:isolation & purification; D004789:Enzyme Activation; D004912:Erythrocytes / Q000201:enzymology*; D006418:Heme / Q000096:biosynthesis*; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D014575:Uroporphyrinogen Decarboxylase / Q000737:chemistry* / Q000302:isolation & purification
D006418:Heme; D003304:Coproporphyrinogen Oxidase; D014575:Uroporphyrinogen Decarboxylase