Advertisement
JBC

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


     


This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Nielsen, F. S.
Right arrow Articles by Jensen, K. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nielsen, F. S.
Right arrow Articles by Jensen, K. F.
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?

Volume 271, Number 46, Issue of November 15, 1996 pp. 29359-29365
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

The B Form of Dihydroorotate Dehydrogenase from Lactococcus lactis Consists of Two Different Subunits, Encoded by the pyrDb and pyrK Genes, and Contains FMN, FAD, and [FeS] Redox Centers

(Received for publication, March 5, 1996, and in revised form, August 19, 1996)

Finn Stausholm Nielsen Dagger § , Paal Skytt Andersen and Kaj Frank Jensen Dagger

From the Dagger  Center for Enzyme Research, Institute of Molecular Biology, University of Copenhagen, Sølvgade 83H, DK-1307 Copenhagen K, Denmark and the  Biotechnological Institute, The Technical University of Denmark, DK-2800 Lyngby, Denmark and the § Department of Biochemistry, University of Illinois, Urbana, Illinois 61801

The B form of dihydroorotate dehydrogenase from Lactococcus lactis (DHOdehase B) is encoded by the pyrDb gene. However, recent genetic evidence has revealed that a co-transcribed gene, pyrK, is needed to achieve the proper physiological function of the enzyme. We have purified DHOdehase B from two strains of Escherichia coli, which harbored either the pyrDb gene or both the pyrDb and the pyrK genes of L. lactis on multicopy plasmids. The enzyme encoded by pyrDb alone (herein called the delta -enzyme) was a bright yellow, dimeric protein that contained one molecule of tightly bound FMN per subunit. The delta -enzyme exhibited dihydroorotate dehydrogenase activity with dichloroindophenol, potassium hexacyanoferrate(III), and molecular oxygen as electron acceptors but could not use NAD+. The DHOdehase B purified from the E. coli strain that carried both the pyrDb and pyrK genes on a multicopy plasmid (herein called the delta kappa -enzyme) was quite different, since it was formed as a complex of equal amounts of the two polypeptides, i.e. two PyrDB and two PyrK subunits. The delta kappa -enzyme was orange-brown and contained 2 mol of FAD, 2 mol of FMN, and 2 mol of [2Fe-2S] redox clusters per mol of native protein as tightly bound prosthetic groups. The delta kappa -enzyme was able to use NAD+ as well as dichloroindophenol, potassium hexacyanoferrate(III), and to some extent molecular oxygen as electron acceptors for the conversion of dihydroorotate to orotate, and it was a considerably more efficient catalyst than the purified delta -enzyme. Based on these results and on analysis of published sequences, we propose that the architecture of the delta kappa -enzyme is representative for the dihydroorotate dehydrogenases from Gram-positive bacteria.


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
Appl. Environ. Microbiol.Home page
S. Kawasaki, T. Satoh, M. Todoroki, and Y. Niimura
b-Type Dihydroorotate Dehydrogenase Is Purified as a H2O2-Forming NADH Oxidase from Bifidobacterium bifidum
Appl. Envir. Microbiol., February 1, 2009; 75(3): 629 - 636.
[Abstract] [Full Text] [PDF]


Home page
J. Bacteriol.Home page
M.-O. Park, T. Mizutani, and P. R. Jones
Glyceraldehyde-3-Phosphate Ferredoxin Oxidoreductase from Methanococcus maripaludis
J. Bacteriol., October 15, 2007; 189(20): 7281 - 7289.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
J. P. Combe, J. Basran, P. Hothi, D. Leys, S. E. J. Rigby, A. W. Munro, and N. S. Scrutton
Lys-D48 Is Required for Charge Stabilization, Rapid Flavin Reduction, and Internal Electron Transfer in the Catalytic Cycle of Dihydroorotate Dehydrogenase B of Lactococcus lactis
J. Biol. Chem., June 30, 2006; 281(26): 17977 - 17988.
[Abstract] [Full Text] [PDF]


Home page
Clin. Chem.Home page
A. B.P. van Kuilenburg, H. van Lenthe, M. Loffler, and A. H. van Gennip
Analysis of Pyrimidine Synthesis "de Novo" Intermediates in Urine and Dried Urine Filter- Paper Strips with HPLC-Electrospray Tandem Mass Spectrometry
Clin. Chem., November 1, 2004; 50(11): 2117 - 2124.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
S. Norager, S. Arent, O. Bjornberg, M. Ottosen, L. L. Leggio, K. F. Jensen, and S. Larsen
Lactococcus lactis Dihydroorotate Dehydrogenase A Mutants Reveal Important Facets of the Enzymatic Function
J. Biol. Chem., August 1, 2003; 278(31): 28812 - 28822.
[Abstract] [Full Text] [PDF]


Home page
J. Bacteriol.Home page
M. Fernandez, M. Kleerebezem, O. P. Kuipers, R. J. Siezen, and R. van Kranenburg
Regulation of the metC-cysK Operon, Involved in Sulfur Metabolism in Lactococcus lactis
J. Bacteriol., January 1, 2002; 184(1): 82 - 90.
[Abstract] [Full Text] [PDF]


Home page
J. Bacteriol.Home page
R. Rebeil, Y. Sun, L. Chooback, M. Pedraza-Reyes, C. Kinsland, T. P. Begley, and W. L. Nicholson
Spore Photoproduct Lyase from Bacillus subtilis Spores Is a Novel Iron-Sulfur DNA Repair Enzyme Which Shares Features with Proteins such as Class III Anaerobic Ribonucleotide Reductases and Pyruvate-Formate Lyases
J. Bacteriol., September 15, 1998; 180(18): 4879 - 4885.
[Abstract] [Full Text]




HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement