A 49-kDa mini-lipoxygenase from Anabaena sp. PCC 7120 retains catalytically complete functionality

The Journal of Biological Chemistry
Yuxiang ZhengAlan R Brash

Abstract

Anabaena sp. PCC 7120 is one of the few prokaryotes harboring a lipoxygenase (LOX) gene. The sequence resides in an open reading frame encoding a fusion protein of a catalase-like hemoprotein with an unusually short LOX (approximately 49 kDa) at the C terminus. The recombinant mini-LOX contains a non-heme iron in the active site and is highly active with linoleic and alpha-linolenic acids (which occur naturally in Anabaena) giving the respective 9R-hydroperoxides, the mirror image of the 9S-LOX products of plants. Using stereospecifically labeled [11-(3)H]linoleic acids we show that reaction is catalyzed via a typical antarafacial relationship of initial hydrogen abstraction and oxygenation. The mini-LOX oxygenated C16/C18:2-phosphatidylcholine with 9R specificity, suggesting a "tail first" mode of fatty acid binding. Site-directed mutagenesis of an active site Ala (Ala215), typically conserved as Gly in R-LOX, revealed that substitution with Gly retained 9R specificity, whereas the larger Val substitution switched oxygenation to 13S, implying that Ala215 represents the functional equivalent of the Gly in other R-LOX. Metabolism studies using a synthetic fatty acid with extended double bond conjugation, 9E,11Z,14Z-20:3omega6, s...Continue Reading

References

Jan 30, 1974·Journal of Chromatography·R H Greeley
May 15, 1980·Analytical Biochemistry·J M BoeynaemsW C Hubbard
Oct 31, 1995·Biochemistry·M H Glickman, Judith P Klinman
Aug 20, 1996·Biochemistry·W MinorB Axelrod
Jun 17, 1998·Proceedings of the National Academy of Sciences of the United States of America·William BoeglinAlan R Brash
Aug 14, 1999·The Journal of Biological Chemistry·Alan R Brash
Sep 14, 2000·The Journal of Biological Chemistry·T HammarbergO Rådmark
Oct 24, 2000·The Journal of Biological Chemistry·X S Chen, C D Funk
Dec 12, 2001·Microbiology·H Porta, M Rocha-Sosa
Sep 12, 2002·Annual Review of Plant Biology·Ivo Feussner, C Wasternack
Nov 16, 2002·Prostaglandins & Other Lipid Mediators·Claus Schneider, Alan R Brash
Nov 16, 2002·Prostaglandins & Other Lipid Mediators·Colin D FunkLei Zhao
Nov 16, 2002·Prostaglandins & Other Lipid Mediators·Ernst H Oliw
Aug 1, 1959·Canadian Journal of Biochemistry and Physiology·E G BLIGH, W J DYER
Oct 1, 2003·Biochemistry·Michael J Knapp, Judith P Klinman
Dec 10, 2003·Current Opinion in Microbiology·James W Golden, Ho-Sung Yoon
Feb 10, 2004·Proceedings of the National Academy of Sciences of the United States of America·Russell E VanceJohn J Mekalanos
Aug 18, 2004·Science·Michael P LesserPaul G Falkowski
Oct 22, 2004·Proceedings of the National Academy of Sciences of the United States of America·Gianguido Coffa, Alan R Brash
Dec 31, 2004·Proceedings of the National Academy of Sciences of the United States of America·Michael L OldhamMarcia E Newcomer
Aug 23, 2005·Biochemical and Biophysical Research Communications·Gianguido CoffaAlan R Brash
Sep 20, 2005·Biochemical and Biophysical Research Communications·Hartmut KuhnMatthias Walther
Apr 7, 2007·Phytochemistry·Imke Lang, Ivo Feussner
May 26, 2007·Chemistry & Biology·Claus SchneiderAlan R Brash
Nov 21, 2007·Proceedings of the National Academy of Sciences of the United States of America·Claus SchneiderAlan R Brash

Citations

Nov 28, 2008·Lipids·Alexandra-Zoi AndreouIvo Feussner
Feb 3, 2015·Critical Reviews in Biotechnology·Ruud HeshofJohanna Buchert
Mar 2, 2010·Chemistry and Physics of Lipids·I Martin-ArjolAngeles Manresa
Sep 22, 2009·Phytochemistry·Alexandra Andreou, Ivo Feussner
Jan 27, 2009·Plant Physiology and Biochemistry : PPB·Alina MosblechIngo Heilmann
Apr 29, 2008·Phytochemistry·Alexandra-Zoi AndreouIvo Feussner
Apr 28, 2012·Biotechnology Advances·Young-Chul Joo, Deok-Kun Oh
Dec 2, 2014·Progress in Lipid Research·Thomas HornDagmar Heydeck
Aug 13, 2013·ChemSusChem·Konrad B OtteBernhard Hauer
Mar 13, 2010·The Journal of Biological Chemistry·Alexandra AndreouIvo Feussner
Nov 28, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ivana HolkováPeter Mikuš

Related Concepts

Bacterial Proteins
Catalysis
Lipoxygenase-2
Proteins, Recombinant DNA
Mutagenesis, Site-Directed
Unassigned Reading Frames
Anabaena
Tertiary Protein Structure
alpha-Linolenic Acid, Tin(2+) Salt
Evolution, Molecular

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.