Iron content of human 5-lipoxygenase, effects of mutations regarding conserved histidine residues.

Recombinant human 5-lipoxygenase was expressed in Escherichia coli and purified to more than 95% homogeneity by ammonium sulfate precipitation and agarose-ATP column chromatography. The specific activity of the purified enzyme was 21-28 mumol/mg, as assessed by the generation of 5-hydro(pero)xyeicosatetraenoic acid. The iron content was analyzed by graphite furnace atomic absorption spectrophotometry for six preparations of the enzyme. The average value of the iron content was 0.86 mol/mol (iron/protein) with a range of 0.74-1.15 mol/mol. All lipoxygenases that have been sequenced contain 6 conserved histidine residues. Mutants of 5-lipoxygenase, with substitutions of these 6 conserved histidines, were purified and analyzed. Mutants H372Q and H550Q had no detectable enzyme activity and were also practically devoid of iron. Three mutants regarding His367 (H367Q, H367N, and H367S) were all inactive but had partial iron contents (0.5, 0.2, and 0.5 mol/mol, respectively). Finally, the mutated proteins H362Q, H390Q, and H399Q displayed reduced enzyme activity but contained similar amounts of iron as non-mutated 5-lipoxygenase. We conclude that histidines 372 and 550 constitute two of the iron ligands in 5-lipoxygenase. Also His367 is necessary for the enzyme activity, but this residue is not crucial for binding of iron.

• Publication year

  1993

• Venue

  Journal of Biological Chemistry

• Publication date

  1993-02-05

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(18)53808-4PMID 8428931
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Mutagenesis studies on the amino acid residues involved in the iron-binding and the activity of human 5-lipoxygenase.

  1992cited by this paper
• Mutagenesis of some conserved residues in human 5-lipoxygenase: effects on enzyme activity.

  1992cited by this paper
• cDNA cloning of rice lipoxygenase L-2 and characterization using an active enzyme expressed from the cDNA in Escherichia coli.

  1992cited by this paper
• Conserved histidine residues in soybean lipoxygenase: functional consequences of their replacement.

  1992cited by this paper
• Recent investigations into the lipoxygenase pathway of plants.

  1991cited by this paper
• Nucleotide sequences of a soybean lipoxygenase gene and the short intergenic region between an upstream lipoxygenase gene

  1991cited by this paper
• Characterization of the activity of purified recombinant human 5-lipoxygenase in the absence and presence of leukocyte factors.

  1991cited by this paper
• Porcine leukocyte 5‐ and 12‐lipoxygenases are iron enzymes

  1991cited by this paper
• Analysis of non-heme iron in arachidonate 12-lipoxygenase of porcine leukocytes.

  1991cited by this paper
• Human 5-lipoxygenase contains an essential iron.

  1991cited by this paper
• Evaluation of the role of conserved His and Met residues among lipoxygenases by site-directed mutagenesis of recombinant human 5-lipoxygenase.

  1991cited by this paper
• The initial characterization of the iron environment in lipoxygenase by Mössbauer spectroscopy.

  1990cited by this paper
• Molecular cloning and expression of human arachidonate 12-lipoxygenase.

  1990cited by this paper
• Alpha-secondary isotope effects in the lipoxygenase reaction.

  1989cited by this paper
• Cloning of the cDNA for human 5-lipoxygenase.

  1988cited by this paper
• Iron environment in soybean lipoxygenase-1.

  1988cited by this paper
• Leukotrienes and lipoxins: structures, biosynthesis, and biological effects.

  1987cited by this paper
• Arachidonate 12-lipoxygenase purified from porcine leukocytes by immunoaffinity chromatography and its reactivity with hydroperoxyeicosatetraenoic acids.

  1986cited by this paper
• On the nature of the 5-lipoxygenase reaction in human leukocytes: enzyme purification and requirement for multiple stimulatory factors.

  1985cited by this paper
• Studies on arachidonate 5-lipoxygenase of rat basophilic leukemia cells.

  1984cited by this paper
• Lipoxygenase from rabbit reticulocytes: iron content, amino acid composition and C-terminal heterogeneity.

  1983cited by this paper
• Leukotrienes: mediators of immediate hypersensitivity reactions and inflammation.

  1983cited by this paper
• Iron, an essential component of lipoxygenase.

  1974cited by this paper
• Soya-bean lipoxygenase: an iron-containing dioxygenase.

  1973cited by this paper
• Soyabean lipoxygenase: an iron-containing enzyme.

  1973cited by this paper

• A site-moiety map and virtual screening approach for discovery of novel 5-LOX inhibitors

  2020cites this paper
• Molecular dynamics study on the Apo‐ and Holo‐forms of 5‐lipoxygenase

  2018cites this paper
• Biosynthesis of leukotriene B4.

  2017cites this paper
• Identification and Characterization of a New Protein Isoform of Human 5-Lipoxygenase

  2016cites this paper
• 5-Lipoxygenase, a key enzyme for leukotriene biosynthesis in health and disease.

  2015cites this paper
• Lipoxygenase : a game-changing enzyme

  2015cites this paper
• Electrophilic Fatty Acid Species Inhibit 5-Lipoxygenase and Attenuate Sepsis-Induced Pulmonary Inflammation

  2014cites this paper
• Methods for identifying lipoxygenase producing microorganisms on agar plates

  2012cites this paper
• Lipoxygenase and leukotriene pathways: biochemistry, biology, and roles in disease.

  2011cites this paper
• Production of 7,10-dihydroxy-8(E)-octadecenoic acid from olive oil by Pseudomonas aeruginosa PR3

  2011cites this paper
• Bioassay-guided identification of an anti-inflammatory prenylated acylphloroglucinol from Melicope ptelefolia and molecular insights into its interaction with 5-lipoxygenase.

  2011cites this paper
• Thermostable lipoxygenase is a key enzyme in the conversion of linoleic acid to trihydroxy-octadecenoic acid by Pseudomonas aeruginosa PR3

  2010cites this paper
• Regioselective Copper-Mediated Organometallic Ring Openings of Acylnitroso Cycloadducts and the Development of 5-Lipoxygenase Inhibitors

  2009cites this paper
• Structural basis for pH-dependent alterations of reaction specificity of vertebrate lipoxygenase isoforms.

  2009cites this paper
• Computational analysis of R and S isoforms of 12-lipoxygenases: homology modeling and docking studies.

  2009cites this paper
• Homology modeling of 5-lipoxygenase and hints for better inhibitor design

  2008cites this paper
• Apoptosis of cultured astrocytes induced by the copper and neocuproine complex through oxidative stress and JNK activation.

  2008cites this paper
• Structural insights into human 5-lipoxygenase inhibition: combined ligand-based and target-based approach.

  2006cites this paper
• Stored iron and vascular reactivity.

  2005cites this paper
• Mutation analysis of the human 5-lipoxygenase C-terminus: support for a stabilizing C-terminal loop.

  2005cites this paper
• Structural organization of the regulatory domain of human 5-lipoxygenase.

  2005cites this paper
• Key Role of Conserved Histidines in Recombinant Mouse β-Carotene 15,15′-Monooxygenase-1 Activity*

  2005cites this paper
• 5-lipoxygenase inhibitors with histamine H(1) receptor antagonist activity.

  2004cites this paper
• Modulation of human 5-lipoxygenase activity by membrane lipids.

  2004cites this paper
• Inhibitory effects of brown algal phlorotannins on secretory phospholipase A2s, lipoxygenases and cyclooxygenases

  2004cites this paper
• Regulation of enzymatic lipid peroxidation: the interplay of peroxidizing and peroxide reducing enzymes.

  2002cites this paper
• Enzymes and receptors in the leukotriene cascade

  2002cites this paper
• Effect of metal ions on the production of isomeric 9,10,13 (9,12,13)-trihydroxy-11E (10E)-octadecenoic acid from linoleic acid by Pseudomonas aeruginosa PR3

  2002cites this paper
• Structural characterization of the catalytic domain of the human 5-lipoxygenase enzyme

  2002cites this paper
• Arachidonate 5-lipoxygenase.

  2002cites this paper
• Effect of metal ions on the production of isomeric 9 , 10 , 13 ( 9 , 12 , 13 )-trihydroxy-11 E ( 10 E )-octadecenoic acid from linoleic acid by Pseudomonas aeruginosa PR 3

  2002cites this paper
• Oxidative stress and c-Jun-amino-terminal kinase activation involved in apoptosis of primary astrocytes induced by disulfiram-Cu(2+) complex.

  2001cites this paper
• EPR investigation of the active site of recombinant human 5-lipoxygenase: inhibition by selenide.

  2001cites this paper
• Structure-Function Investigation of the Interaction of 1- and 2-Substituted 3-Hydroxypyridin-4-ones with 5-Lipoxygenase and Ribonucleotide Reductase*

  2001cites this paper
• Mutagenesis and modelling of linoleate-binding to pea seed lipoxygenase.

  2001cites this paper
• Leukotriene A4 hydrolase : Identification of amino acid residues involved in catalyses and substrate-mediated inactivation

  2001cites this paper
• Oxidative stress and c-Jun-amino-terminal kinase activation involved in apoptosis of primary astrocytes induced by disulfiram-Cu2+ complex

  2001cites this paper
• Identification of amino acid residues involved in catalyses and substrate-mediated inactivation

  2001cites this paper
• A simple and efficient method for hemoglobin removal from mammalian tissue cytosol by zinc sulfate and its application to the study of lipoxygenase.

  2000cites this paper
• Glutathione peroxidase-1 but not -4 is involved in the regulation of cellular 5-lipoxygenase activity in monocytic cells.

  2000cites this paper
• 5-Lipoxygenase and leukotrienes. Transgenic mouse and nuclear targeting studies.

  2000cites this paper
• Death signaling pathway induced by pyrrolidine dithiocarbamate‐Cu2+ complex in the cultured rat cortical astrocytes

  2000cites this paper
• Analysis of a nucleotide-binding site of 5-lipoxygenase by affinity labelling: binding characteristics and amino acid sequences.

  2000cites this paper
• 5-Lipoxygenase: a target for antiinflammatory drugs revisited.

  1999cites this paper
• Mutagenesis Studies of Mammalian Lipoxygenases

  1999cites this paper
• The biology of 5-lipoxygenase: function, structure, and regulatory mechanisms.

  1999cites this paper
• Nonredox 5-lipoxygenase inhibitors require glutathione peroxidase for efficient inhibition of 5-lipoxygenase activity.

  1998cites this paper
• Determinants of 5-Lipoxygenase Nuclear Localization Using Green Fluorescent Protein/5-Lipoxygenase Fusion Proteins*

  1998cites this paper
• Structure of soybean lipoxygenase L3 and a comparison with its L1 isoenzyme

  1997cites this paper
• Molecular and catalytic properties of mammalian lipoxygenases compared with soybean lipoxygenase-1.

  1997cites this paper
• Lipoxygenases of Mice and Men

  1996cites this paper
• Selenite incubated with NADPH and mammalian thioredoxin reductase yields selenide, which inhibits lipoxygenase and changes the electron spin resonance spectrum of the active site iron.

  1996cites this paper
• The Environment of the Lipoxygenase Iron Binding Site Explored with Novel Hydroxypyridinone Iron Chelators (*)

  1996cites this paper
• Exploring the Structure and Function of Mammalian Lipoxygenases by Site-Directed Mutagenesis

  1996cites this paper
• Dithiocarbamates Induce Apoptosis in Thymocytes by Raising the Intracellular Level of Redox-active Copper (*)

  1995cites this paper
• 5-lipoxygenase: structure and stability of recombinant enzyme, regulation in Mono Mac 6 cells.

  1995cites this paper
• Mutations at the C-terminal isoleucine and other potential iron ligands of 5-lipoxygenase.

  1995cites this paper
• Arachidonate 5-lipoxygenase.

  1995cites this paper
• Sequential induction of 5-lipoxygenase gene expression and activity in Mono Mac 6 cells by transforming growth factor beta and 1,25-dihydroxyvitamin D3.

  1995cites this paper
• Specfficity of expression and effects of eicosanoid

  1995cites this paper
• Specificity of expression and effects of eicosanoid mediators in normal physiology and human diseases 1

  1995cites this paper
• Stabilization of purified human 5-lipoxygenase with glutathione peroxidase and superoxide dismutase.

  1994cites this paper
• Interfacial kinetic reaction of human 5-lipoxygenase.

  1994cites this paper
• 5-Lipoxygenase.

  1994cites this paper
• 5-Lipoxygenase: enzyme expression and regulation of activity.

  1994cites this paper
• Site-directed mutagenesis studies on the iron-binding domain and the determinant for the substrate oxygenation site of porcine leukocyte arachidonate 12-lipoxygenase.

  1994cites this paper
• Crystallographic determination of the active site iron and its ligands in soybean lipoxygenase L-1.

  1993cites this paper
• Characterization of the non-heme iron center of human 5-lipoxygenase by electron paramagnetic resonance, fluorescence, and ultraviolet-visible spectroscopy: redox cycling between ferrous and ferric states.

  1993cites this paper