Lipoxin and resolvin biosynthesis is dependent on 5-lipoxygenase activating protein.
Resolution of acute inflammation is an active process coordinated by proresolving lipid mediators (SPMs) such as lipoxins (LXs) and resolvins (Rvs), which are formed by the concerted action of 2 lipoxygenases (LOs). Because the exact molecular mechanisms of SPM biosynthesis are not completely understood, we aimed to investigate LX and D-type Rv formation in human leukocytes and HEK293T cells overexpressing leukotriene (LT) pathway enzymes. Activity assays in precursor (15-hydroxyeicosatetraenoic acids, 17-HDoHE)-treated granulocytes [polymorphonuclear leukocytes (PMNLs)] showed a strict dependence of LXA4/RvD1 biosynthesis on cell integrity, and incubation with recombinant human 5-LO did not lead to LX or Rv formation. Pharmacologic inhibition of 5-LO activating protein (FLAP) by MK-886 inhibited LXA4/RvD1 biosynthesis in precursor-treated PMNLs (drug concentration causing 50% inhibition ∼ 0.3/0.2 µM), as did knockdown of the enzyme in MM6 cells, and precursor-treated HEK293T overexpressing 5-LO produced high amounts of LXA4 only in the presence of FLAP. In addition, inhibition of cytosolic phospholipase A2α (cPLA2α) interfered with LXA4/RvD1 formation from exogenous precursors in PMNLs. Furthermore, inhibition of the LT synthases LTA4 hydrolase and LTC4 synthase in PMNL/platelet coincubations augmented LXA4 levels. These findings show that several enzymes known to be involved in the biosynthesis of proinflammatory LTs, such as FLAP and cPLA2α, also contribute to LX and Rv formation.
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Christoph
C
Lehmann
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Julia
J
Homann
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Ann-Katrin
AK
Ball
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
René
R
Blöcher
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Thea K
TK
Kleinschmidt
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Devaraj
D
Basavarajappa
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Carlo
C
Angioni
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Nerea
N
Ferreirós
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Ann-Kathrin
AK
Häfner
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Olof
O
Rådmark
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Ewgenij
E
Proschak
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Jesper Z
JZ
Haeggström
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Gerd
G
Geisslinger
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Michael J
MJ
Parnham
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden.
Dieter
D
Steinhilber
*Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Translational Medicine and Pharmacology, Institute of Clinical Pharmacology, Pharmazentrum Frankfurt, and Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany; and Department of Medical Biochemistry and Biophysics, Division of Chemistry 2, Karolinska Institutet, Stockholm, Sweden kahnt@pharmchem.uni-frankfurt.de.
Astrid Stefanie
AS
Kahnt
D058927:5-Lipoxygenase-Activating Proteins / Q000378:metabolism*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D045744:Cell Line, Tumor; D003600:Cytosol / Q000201:enzymology; D004281:Docosahexaenoic Acids / Q000096:biosynthesis*; D054513:Group IV Phospholipases A2 / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D044045:Lipoxins / Q000096:biosynthesis*; D008264:Macrophages / Q000201:enzymology / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects; D011994:Recombinant Proteins / Q000378:metabolism; D002722:Chlorobenzenes
D058927:5-Lipoxygenase-Activating Proteins; D001094:Arachidonate 5-Lipoxygenase; D004281:Docosahexaenoic Acids; D054513:Group IV Phospholipases A2; D007211:Indoles; D044045:Lipoxins; D011994:Recombinant Proteins; C040527:lipoxin A4; C518399:resolvin D1; C060893:MK-886; D002722:Chlorobenzenes
LTA4 hydrolase; LTC4 synthase; cytosolic phospholipase A2α; resolution of inflammation