Oxygen-dependent coproporphyrinogen-III oxidase from Escherichia coli: one-step purification and biochemical characterisation.

Abstract

Coproporphyrinogen-III oxidase (CPO) catalyses the conversion of coproporphyrinogen-III to protoporphyrinogen-IX in the haem biosynthetic pathway, and its deficient activity is associated with human hereditary coproporphyria. The 47% sequence identity between the oxygen-dependent CPO from Escherichia coli and its human counterpart makes the bacterial enzyme a good model system for structural studies of this disease. Therefore, we overexpressed and purified to homogeneity the oxygen-dependent CPO from E. coli and its selenomethionine derivative fused with a His(6)-tag. Both preparations showed a specific activity of 37500 U mg(-1), had a subunit molecular mass of 35 kDa and behaved as a compact shaped dimer. First crystallisation trials produced plate-shaped diffracting crystals.

• Authors: Sofia Macieira, Berta M Martins, Robert Huber
• Journal: FEMS microbiology letters (FEMS Microbiol Lett) Vol. 226 Issue 1 Pg. 31-7 (Sep 12 2003) ISSN: 0378-1097 [Print] England
• PMID: 13129604 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Coproporphyrinogens
• Protein Subunits
• Protoporphyrins
• Recombinant Proteins
• coproporphyrinogen III
• protoporphyrin IX
• Coproporphyrinogen Oxidase

Topics

• Amino Acid Sequence
• Coproporphyrinogen Oxidase (chemistry, genetics, isolation & purification, metabolism)
• Coproporphyrinogens (metabolism)
• Crystallization
• Crystallography, X-Ray
• Electrophoresis, Polyacrylamide Gel
• Escherichia coli (enzymology)
• Gene Expression Regulation, Bacterial
• Molecular Sequence Data
• Protein Subunits (chemistry)
• Protoporphyrins (metabolism)
• Recombinant Proteins (genetics, isolation & purification, metabolism)
• Spectrum Analysis