Lipoxygenase

Lipoxygenase

Structure of rabbit reticulocyte 15S-lipoxygenase.[1]
Identifiers
Symbol Lipoxygenase
Pfam PF00305
InterPro IPR013819
PROSITE PDOC00077
SCOP 2sbl
SUPERFAMILY 2sbl
OPM superfamily 87
OPM protein 2p0m

Lipoxygenases (EC 1.13.11.-) are a family of iron-containing enzymes most of which catalyze the dioxygenation of polyunsaturated fatty acids in lipids containing a cis,cis-1,4- pentadiene structure as shown in the following reaction:

Fatty acid + O2 = fatty acid hydroperoxide

The lipoxygenases are related to each other based upon their similar genetic structure and dioxygenation activity. However, one lipoxygenase, ALOXE3, while having a lipoxygenase genetic structure, possesses relatively little dioxygenation activity; rather its primary activity appears to be as an isomerase that catalyzes the conversion of hydroperoxy unsaturated fatty acids to their 1,5-epoxide,hydroxyl derivatives.

Lipoxygenases are found in plants, animals and fungi. Products of lipoxygenases are involved in diverse cell functions.

Biological function and classification

These enzymes are most common in plants where they may be involved in a number of diverse aspects of plant physiology including growth and development, pest resistance, and senescence or responses to wounding.[2] In mammals a number of lipoxygenases isozymes are involved in the metabolism of eicosanoids (such as prostaglandins, leukotrienes and nonclassic eicosanoids).[3] Sequence data is available for the following lipoxygenases:

Plant lipoxygenases

Plants express a variety of cytosolic lipoxygenases (EC 1.13.11.12InterPro: IPR001246) as well as what seems to be a chloroplast isozyme.[4]

Human lipoxygenases

With the exception of the 5-LOX gene which is located on chromosome 10q11.2, all six human LOX genes are located on chromosome 17.p13 and code for a single chain protein of 75–81 kiloDaltons and consisting of 662–711 amino acids. Mammalian LOX genes contain 14 (ALOX5, ALOX12, ALOX15, ALOX15B) or 15 (ALOX12B, ALOXE3) exons with exon/intron boundaries at highly conserved position.[5][6] The 6 human lipoxygenases along with some of the major products that they make as well as some their associations with genetic diseases are as follows:[5][7]

Two lipoxygenases may act in series to make di-hydroxy or tri-hydroxy products that have activities quite different than either lipoxyenases' products. This serial metabolism may occur in different cell types that express only one of the two lipoxygenases in a process termed transcellular metabolism. For example, ALOX5 and ALOX15 or, alternatively, ALOX5 and ALOX12 can act serially to metabolize arachidonic acid into lipoxins (see 15-hydroxyicosatetraenoic acid#Further metabolism of 15(S)-HpETE, 15(S)-HETE, 15(R)-HpETE, 15(R)-HETE, and 15-oxo-ETE and lipoxin#Biosynthesis) while ALOX15 and possibly ALOX15B can act with ALOX5 to metabolize eicosapentaenoic acid to resolvin D's (see resolvin#Production).

Mouse lipoxygenases

The mouse is a common model to examine lipoxygenase function. However, there are some key differences between the lipoxygenases between mice and men that make extrapolations from mice studies to humans difficult. In contrast to the 6 functional lipoxygenases in humans, mice have 7 functional lipoxygenases and some of the latter have different metabolic activities than their human orthologs.[10][12][13] In particular, mouse Alox15, unlike human ALOX15, metabolizes arachidonic acid mainly to 12-HpETE and mouse Alox15b, in contrast to human ALOX15b, is primarily an 8-lipoxygenase, metabolizing arachdionic acid to 8-HpETE; there is no comparable 8-HpETE-forming lipoxygenase in humans.[14]

Rabbit 15-lipoxygenase (blue) with inhibitor (yellow) bound in the active site

3D structure

There are several lipoxygenase structures known including: soybean lipoxygenase L1 and L3, coral 8-lipoxygenase, human 5-lipoxygenase, rabbit 15-lipoxygenase and porcine leukocyte 12-lipoxygenase catalytic domain. The protein consists of a small N-terminal PLAT domain and a major C-terminal catalytic domain (see Pfam link in this article), which contains the active site. In both plant and mammalian enzymes, the N-terminal domain contains an eight-stranded antiparallel β-barrel, but in the soybean lipoxygenases this domain is significantly larger than in the rabbit enzyme. The plant lipoxygenases can be enzymatically cleaved into two fragments which stay tightly associated while the enzyme remains active; separation of the two domains leads to loss of catalytic activity. The C-terminal (catalytic) domain consists of 18-22 helices and one (in rabbit enzyme) or two (in soybean enzymes) antiparallel β-sheets at the opposite end from the N-terminal β-barrel.

Active site

The iron atom in lipoxygenases is bound by four ligands, three of which are histidine residues.[15] Six histidines are conserved in all lipoxygenase sequences, five of them are found clustered in a stretch of 40 amino acids. This region contains two of the three zinc-ligands; the other histidines have been shown[16] to be important for the activity of lipoxygenases.

The two long central helices cross at the active site; both helices include internal stretches of π-helix that provide three histidine (His) ligands to the active site iron. Two cavities in the major domain of soybean lipoxygenase-1 (cavities I and II) extend from the surface to the active site. The funnel-shaped cavity I may function as a dioxygen channel; the long narrow cavity II is presumably a substrate pocket. The more compact mammalian enzyme contains only one boot-shaped cavity (cavity II). In soybean lipoxygenase-3 there is a third cavity which runs from the iron site to the interface of the β-barrel and catalytic domains. Cavity III, the iron site and cavity II form a continuous passage throughout the protein molecule.

The active site iron is coordinated by Nε of three conserved His residues and one oxygen of the C-terminal carboxyl group. In addition, in soybean enzymes the side chain oxygen of asparagine is weakly associated with the iron. In rabbit lipoxygenase, this Asn residue is replaced with His which coordinates the iron via Nδ atom. Thus, the coordination number of iron is either five or six, with a hydroxyl or water ligand to a hexacoordinate iron.

Details about the active site feature of lipoxygenase were revealed in the structure of porcine leukocyte 12-lipoxygenase catalytic domain complex [15][17] In the 3D structure, the substrate analog inhibitor occupied a U-shaped channel open adjacent to the iron site. This channel could accommodate arachidonic acid without much computation, defining the substrate binding details for the lipoxygenase reaction. In addition, a plausible access channel, which intercepts the substrate binding channel and extended to the protein surface could be counted for the oxygen path.

Biochemical classification

EC 1.13.11.12 lipoxygenase (linoleate:oxygen 13-oxidoreductase) linoleate + O2 = (9Z,11E,13S)-13-hydroperoxyoctadeca-9,11-dienoate
EC 1.13.11.31 arachidonate 12-lipoxygenase (arachidonate:oxygen 12-oxidoreductase) arachidonate + O2 = (5Z,8Z,10E,12S,14Z)-12-hydroperoxyicosa-5,8,10,14-tetraenoate
EC 1.13.11.33 arachidonate 15-lipoxygenase (arachidonate:oxygen 15-oxidoreductase) arachidonate + O2 = (5Z,8Z,11Z,13E,15S)-15-hydroperoxyicosa-5,8,11,13-tetraenoate
EC 1.13.11.34 arachidonate 5-lipoxygenase (arachidonate:oxygen 5-oxidoreductase) arachidonate + O2 = leukotriene A4 + H2
EC 1.13.11.40 arachidonate 8-lipoxygenase (arachidonate:oxygen 8-oxidoreductase) arachidonate + O2 = (5Z,8R,9E,11Z,14Z)-8-hydroperoxyicosa-5,9,11,14-tetraenoate

Soybean Lipoxygenase 1 exhibits the largest H/D kinetic isotope effect (KIE) on kcat (kH/kD) (81 near room temperature) so far reported for a biological system. Recently, an extremely elevated KIE of 540 to 730 was found in a double mutant Soybean Lipoxygenase 1.[18] Because of the large magnitude of the KIE, Soybean Lipoxygenase 1 has served as the prototype for enzyme-catalyzed hydrogen-tunneling reactions.

Human proteins expressed from the lipoxygenase family include ALOX12, ALOX12B, ALOX15, ALOX15B, ALOX5, and ALOXE3. While humans also possess the ALOX12P2 gene, which is an ortholog of the well-expressed Alox12P gene in mice, the human gene is a pseudogene; consequently, ALOX12P2 protein is not detected in humans.[19]

References

  1. Choi J, Chon JK, Kim S, Shin W (February 2008). "Conformational flexibility in mammalian 15S-lipoxygenase: Reinterpretation of the crystallographic data". Proteins 70 (3): 1023–32. doi:10.1002/prot.21590. PMID 17847087.
  2. Vick BA, Zimmerman DC (1987). "Oxidative systems for the modification of fatty acids" 9: 53–90. doi:10.1016/b978-0-12-675409-4.50009-5.
  3. Needleman P, Turk J, Jakschik BA, Morrison AR, Lefkowith JB (1986). "Arachidonic acid metabolism". Annu. Rev. Biochem. 55: 69–102. doi:10.1146/annurev.bi.55.070186.000441. PMID 3017195.
  4. Tanaka K, Ohta H, Peng YL, Shirano Y, Hibino T, Shibata D (1994). "A novel lipoxygenase from rice. Primary structure and specific expression upon incompatible infection with rice blast fungus". J. Biol. Chem. 269 (5): 3755–3761. PMID 7508918.
  5. 1 2 Biochim Biophys Acta. 2014 Mar;1841(3):390-400. doi: 10.1016/j.bbalip.2013.08.005. Epub 2013 Aug 16. Review. Erratum in: Biochim Biophys Acta. 2014 Dec;1841(12):1767. PMID 23954555
  6. http://www.ncbi.nlm.nih.gov/gene/240
  7. Chem Rev. 2011 Oct 12;111(10):5866-98. doi: 10.1021/cr200246d. Epub 2011 Sep 22. Review. No abstract available. PMID 21936577
  8. https://www.wikigenes.org/e/gene/e/247.html
  9. 1 2 https://www.wikigenes.org/e/gene/e/242.html
  10. 1 2 3 4 Biochim Biophys Acta. 2014 Mar;1841(3):401-8. doi: 10.1016/j.bbalip.2013.08.020. Epub 2013 Sep 7. Review.PMID 24021977
  11. 1 2 https://www.wikigenes.org/e/gene/e/59344.html
  12. Biochim Biophys Acta. 2014 Mar;1841(3):390-400. doi:10.1016/j.bbalip.2013.08.005. Epub 2013 Aug 16. Review. Erratum in: Biochim Biophys Acta. 2014 Dec;1841(12):1767.PMID 23954555
  13. Blood. 2012 Jun 21;119(25):6173-4; author reply 6174-5. doi:10.1182/blood-2012-02-410928. PMID 22730527
  14. Prostaglandins Other Lipid Mediat. 2013 Jul-Aug;104-105:84-92. doi: 10.1016/j.prostaglandins.2012.07.004. Epub 2012 Aug 20. Review.PMID 22951339
  15. 1 2 Boyington JC, Gaffney BJ, Amzel LM (1993). "The three-dimensional structure of an arachidonic acid 15-lipoxygenase". Science 260 (5113): 1482–1486. doi:10.1126/science.8502991. PMID 8502991.
  16. Steczko J, Donoho GP, Clemens JC, Dixon JE, Axelrod B (1992). "Conserved histidine residues in soybean lipoxygenase: functional consequences of their replacement". Biochemistry 31 (16): 4053–4057. doi:10.1021/bi00131a022. PMID 1567851.
  17. Xu, S.; Mueser T.C.; Marnett L.J.; Funk M.O. (2012). "Crystal structure of 12-lipoxygenase catalytic-domain-inhibitor complex identifies a substrate-binding channel for catalysis.". Structure 20 (9): 1490–7. doi:10.1016/j.str.2012.06.003. PMID 22795085.
  18. Hu S1, Sharma SC, Scouras AD, Soudackov AV, Carr CA, Hammes-Schiffer S, Alber T, Klinman JP. (2014). "Extremely elevated room-temperature kinetic isotope effects quantify the critical role of barrier width in enzymatic C-H activation". JACS 136 (23):8157-60.
  19. https://www.wikigenes.org/search.html?search=ALOX12P2

External links

This article incorporates text from the public domain Pfam and InterPro IPR001024

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