Human coproporphyrinogen oxidase is not a metalloprotein.

The Journal of Biological Chemistry
A E Medlock, H A Dailey

Abstract

Coproporphyrinogen oxidase (CPO) (EC 1.3.3.3), the antepenultimate enzyme in the heme biosynthetic pathway, catalyzes the conversion of coproporphyrinogen III to protoporphyrinogen IX. Previously, based upon metal analysis and site-directed mutagenesis of purified recombinant enzyme, it has been suggested that CPO contains and requires copper for activity (Kohno, H., Furukawa, T., Tokunaga, R., Taketani, S., and Yoshinaga, T. (1996) Biochim. Biophys. Acta 1292, 156-162). To examine this putative metal site in human CPO, the cDNA encoding human CPO was engineered into an expression vector with a His6 tag at its amino terminus, and the protein was expressed in Escherichia coli and purified to apparent homogeneity using nickel-nitroliotriacetic acid resin. Activity of the purified protein was monitored by a coupled fluorometric assay that employed purified protoporphyrinogen oxidase to convert protoporphyrinogen to protoporphyrin, thereby allowing the direct fluorescent determination of protoporphyrin IX produced. CPO has an apparent Km of 0.6 microM and an apparent Kcat of 16 min-1 with coproporphyrinogen III as substrate. Metal analysis of the enzyme was carried out via ultraviolet and visible spectroscopy, inductively coupled p...Continue Reading

References

Oct 15, 1978·The Biochemical Journal·B GrandchampY Nordmann
May 1, 1989·European Journal of Biochemistry·M BogardP Labbe
May 2, 1986·European Journal of Biochemistry·J M CamadroP Labbe
Oct 1, 1985·Analytical Biochemistry·P K SmithD C Klenk
Aug 15, 1985·Analytical Biochemistry·P LabbeH Chambon
Jan 1, 1982·Enzyme·B Grandchamp, Y Nordmann
Apr 12, 1994·Proceedings of the National Academy of Sciences of the United States of America·P MartasekB Grandchamp
Oct 1, 1993·Plant Molecular Biology·O MadsenK A Marcker
Jan 4, 1994·Biochimica Et Biophysica Acta·S TaketaniR Tokunaga
Jan 1, 1996·Protein Science : a Publication of the Protein Society·T A Dailey, H A Dailey

❮ Previous
Next ❯

Citations

Apr 23, 2003·Journal of Biochemical and Biophysical Methods·M A JonesT D Lash
Nov 18, 2000·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·E M Sorianello, M B Mazzetti
Mar 23, 2002·Bioorganic & Medicinal Chemistry Letters·Timothy D LashMarjorie A Jones
Sep 24, 2005·Proceedings of the National Academy of Sciences of the United States of America·Dong-Sun LeeC S Raman
Dec 17, 2009·Parasitology International·Viswanathan Arun NagarajGovindarajan Padmanaban
Jan 31, 2014·Chemical Reviews·Joan B BroderickEric M Shepard
Mar 13, 2014·Animal Science Journal = Nihon Chikusan Gakkaihō·Chuanwei ZhengZhonghua Ning
May 28, 2010·Protein Science : a Publication of the Protein Society·Gunhild LayerDirk W Heinz
Dec 1, 2004·Annual Review of Genetics·Arthur R GrossmanChung Soon Im
Sep 17, 2003·The Journal of Biological Chemistry·Daniela BreckauDieter Jahn
Jul 13, 2002·The Journal of Biological Chemistry·Gunhild LayerDieter Jahn
Jun 15, 2004·The Journal of Biological Chemistry·John D PhillipsChristopher P Hill
Apr 1, 2007·EcoSal Plus·Samuel I Beale
Feb 5, 2011·The Journal of Physical Chemistry. B·Pedro J Silva, Maria João Ramos
Mar 4, 2008·Archives of Biochemistry and Biophysics·Ilka U HeinemannDieter Jahn

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.