JBC Oz Biosciences

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M205247200 on July 11, 2002

J. Biol. Chem., Vol. 277, Issue 37, 34136-34142, September 13, 2002
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
277/37/34136    most recent
M205247200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Layer, G.
Right arrow Articles by Jahn, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Layer, G.
Right arrow Articles by Jahn, D.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Oxygen-independent Coproporphyrinogen-III Oxidase HemN from Escherichia coli*

Gunhild LayerDagger , Knut Verfürth§, Esther MahlitzDagger , and Dieter JahnDagger

From the Dagger  Institut für Mikrobiologie, Technische Universität Braunschweig, D-38106 Braunschweig, Germany and the § Institut für Biologie II, Mikrobiologie, Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg im Breisgau, Germany

In bacteria the oxygen-independent coproporphyrinogen-III oxidase catalyzes the oxygen-independent conversion of coproporphyrinogen-III to protoporphyrinogen-IX. The Escherichia coli hemN gene encoding a putative part of this enzyme was overexpressed in E. coli. Anaerobically purified HemN is a monomeric protein with a native Mr = 52,000 ± 5,000. A newly established anaerobic enzyme assay was used to demonstrate for the first time in vitro coproporphyrinogen-III oxidase activity for recombinant purified HemN. The enzyme requires S-adenosyl-L-methionine (SAM), NAD(P)H, and additional cytoplasmatic components for catalysis. An oxygen-sensitive iron-sulfur cluster was identified by absorption spectroscopy and iron analysis. Cysteine residues Cys62, Cys66, and Cys69, which are part of the conserved CXXXCXXC motif found in all HemN proteins, are essential for iron-sulfur cluster formation and enzyme function. Completely conserved residues Tyr56 and His58, localized closely to the cysteine-rich motif, were found to be important for iron-sulfur cluster integrity. Mutation of Gly111 and Gly113, which are part of the potential GGGTP S-adenosyl-L-methionine binding motif, completely abolished enzymatic function. Observed functional properties in combination with a recently published computer-based enzyme classification (Sofia, H. J., Chen, G., Hetzler, B. G., Reyes-Spindola, J. F., and Miller, N. E. (2001) Nucleic Acids Res. 29, 1097-1106) identifies HemN as "Radical SAM enzyme." An appropriate enzymatic mechanism is suggested.

* This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Institut für Mikrobiologie, Technische Universität Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, Germany. Tel.: 49-531-3915801; Fax: 49-531-3915854; E-mail: d.jahn@tu-bs.de.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 MicrobiologyHome page
A. Masoumi, I. U. Heinemann, M. Rohde, M. Koch, M. Jahn, and D. Jahn
Complex formation between protoporphyrinogen IX oxidase and ferrochelatase during haem biosynthesis in Thermosynechococcus elongatus
Microbiology, December 1, 2008; 154(12): 3707 - 3714.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
Y.-L. Chan, T.-H. Chang, C.-L. Liao, and Y.-L. Lin
The Cellular Antiviral Protein Viperin Is Attenuated by Proteasome-Mediated Protein Degradation in Japanese Encephalitis Virus-Infected Cells
J. Virol., November 1, 2008; 82(21): 10455 - 10464.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. J. Brocker, S. Virus, S. Ganskow, P. Heathcote, D. W. Heinz, W.-D. Schubert, D. Jahn, and J. Moser
ATP-driven Reduction by Dark-operative Protochlorophyllide Oxidoreductase from Chlorobium tepidum Mechanistically Resembles Nitrogenase Catalysis
J. Biol. Chem., April 18, 2008; 283(16): 10559 - 10567.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Chandor, O. Berteau, T. Douki, D. Gasparutto, Y. Sanakis, S. Ollagnier-de-Choudens, M. Atta, and M. Fontecave
Dinucleotide Spore Photoproduct, a Minimal Substrate of the DNA Repair Spore Photoproduct Lyase Enzyme from Bacillus subtilis
J. Biol. Chem., September 15, 2006; 281(37): 26922 - 26931.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Layer, A. J. Pierik, M. Trost, S. E. Rigby, H. K. Leech, K. Grage, D. Breckau, I. Astner, L. Jansch, P. Heathcote, et al.
The Substrate Radical of Escherichia coli Oxygen-independent Coproporphyrinogen III Oxidase HemN
J. Biol. Chem., June 9, 2006; 281(23): 15727 - 15734.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Layer, K. Grage, T. Teschner, V. Schunemann, D. Breckau, A. Masoumi, M. Jahn, P. Heathcote, A. X. Trautwein, and D. Jahn
Radical S-Adenosylmethionine Enzyme Coproporphyrinogen III Oxidase HemN: FUNCTIONAL FEATURES OF THE [4Fe-4S] CLUSTER AND THE TWO BOUND S-ADENOSYL-L-METHIONINES
J. Biol. Chem., August 12, 2005; 280(32): 29038 - 29046.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. D. Phillips, F. G. Whitby, C. A. Warby, P. Labbe, C. Yang, J. W. Pflugrath, J. D. Ferrara, H. Robinson, J. P. Kushner, and C. P. Hill
Crystal Structure of the Oxygen-dependant Coproporphyrinogen Oxidase (Hem13p) of Saccharomyces cerevisiae
J. Biol. Chem., September 10, 2004; 279(37): 38960 - 38968.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Nicolet and C. L. Drennan
AdoMet radical proteins--from structure to evolution--alignment of divergent protein sequences reveals strong secondary structure element conservation
Nucleic Acids Res., August 2, 2004; 32(13): 4015 - 4025.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Q. Fang, J. Peng, and T. Dierks
Post-translational Formylglycine Modification of Bacterial Sulfatases by the Radical S-Adenosylmethionine Protein AtsB
J. Biol. Chem., April 9, 2004; 279(15): 14570 - 14578.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Ouchane, A.-S. Steunou, M. Picaud, and C. Astier
Aerobic and Anaerobic Mg-Protoporphyrin Monomethyl Ester Cyclases in Purple Bacteria: A STRATEGY ADOPTED TO BYPASS THE REPRESSIVE OXYGEN CONTROL SYSTEM
J. Biol. Chem., February 20, 2004; 279(8): 6385 - 6394.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
C. Baysse, S. Matthijs, M. Schobert, G. Layer, D. Jahn, and P. Cornelis
Co-ordination of iron acquisition, iron porphyrin chelation and iron-protoporphyrin export via the cytochrome c biogenesis protein CcmC in Pseudomonas fluorescens
Microbiology, December 1, 2003; 149(12): 3543 - 3552.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. Breckau, E. Mahlitz, A. Sauerwald, G. Layer, and D. Jahn
Oxygen-dependent Coproporphyrinogen III Oxidase (HemF) from Escherichia coli Is Stimulated by Manganese
J. Biol. Chem., November 21, 2003; 278(47): 46625 - 46631.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Marquordt, Q. Fang, E. Will, J. Peng, K. von Figura, and T. Dierks
Posttranslational Modification of Serine to Formylglycine in Bacterial Sulfatases. RECOGNITION OF THE MODIFICATION MOTIF BY THE IRON-SULFUR PROTEIN AtsB
J. Biol. Chem., January 17, 2003; 278(4): 2212 - 2218.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.