Literature summary for 1.13.11.61 extracted from

Newie, J.; Neumann, P.; Werner, M.; Mata, R.A.; Ficner, R.; Feussner, I.
Lipoxygenase 2 from Cyanothece sp. controls dioxygen insertion by steric shielding and substrate fixation (2017), Sci. Rep., 7, 2069 .

Search for more literature for 1.13.11.61

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21 Star	Rippkaea orientalis PCC 8801

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type enzyme CspLOX2, sitting drop vapor diffusion method, from well solution of 10% PEG 4000, 0.1 M MES/imidazole pH 6.5, 20% glycerol and 0.02% alcohols (1,6-hexanediol, 1-butanol, 1,2-propanediol, 2-propanol, 1,4-butanediol, and 1,3-propanediol), 10 days, purified recombinant CspLOX2 mutant enzymes by hanging drop vapour diffusion method, from well solution of 12.5-15 PEG 4000, 8-12% glycerol, 0.1 M MES/imidazole pH 6.1, and 0-600 mM NaCl, X-ray diffraction structure determination and analysis at 1.8 A resolution. Crystals of a CspLOX2 substrate complex are not obtained	Rippkaea orientalis PCC 8801

Crystallization (Comment)

Organism

purified recombinant wild-type enzyme CspLOX2, sitting drop vapor diffusion method, from well solution of 10% PEG 4000, 0.1 M MES/imidazole pH 6.5, 20% glycerol and 0.02% alcohols (1,6-hexanediol, 1-butanol, 1,2-propanediol, 2-propanol, 1,4-butanediol, and 1,3-propanediol), 10 days, purified recombinant CspLOX2 mutant enzymes by hanging drop vapour diffusion method, from well solution of 12.5-15 PEG 4000, 8-12% glycerol, 0.1 M MES/imidazole pH 6.1, and 0-600 mM NaCl, X-ray diffraction structure determination and analysis at 1.8 A resolution. Crystals of a CspLOX2 substrate complex are not obtained

Rippkaea orientalis PCC 8801

Protein Variants

Protein Variants	Comment	Organism
G401A	site-directed mutagenesis, the mutaant shows increased formation of 11-hydroperoxyoctadeca-9,12-dienoate compared to the wild-type enzyme	Rippkaea orientalis PCC 8801
I617L	site-directed mutagenesis, the mutaant shows increased formation of 11-hydroperoxyoctadeca-9,12-dienoate compared to the wild-type enzyme	Rippkaea orientalis PCC 8801
additional information	Decreasing the channel size of a 9R-lipoxygenase (CspLOX1) can in turn induce formation of the bis-allylic 11R-hydroperoxide	Rippkaea orientalis PCC 8801

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe3+	non-heme iron, required	Rippkaea orientalis PCC 8801

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
linoleate + O2	Rippkaea orientalis PCC 8801	-	(9Z,11R,12Z)-11-hydroperoxyoctadeca-9,12-dienoate	-	?

Organism

Organism	UniProt	Comment	Textmining
Rippkaea orientalis PCC 8801	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21 Star by nickel affinity chromatography, gel filtration, and ultrafiltration	Rippkaea orientalis PCC 8801

Reaction

Reaction	Comment	Organism	Reaction ID
alpha-linolenate + O2 = (9R,10E,12Z,15Z)-9-hydroperoxyoctadeca-10,12,15-trienoate	the LOX reaction is always catalyzed at a 1Z,4Z-pentadiene system of a polyunsaturated fatty acid. In the initial step, a hydrogen atom is abstracted from the central bis-allylic methylene group by the non-heme iron or in few cases manganese in the active site. During this rate-limiting step, Fe(III) is reduced to Fe(II) in the former case and a substrate radical is formed which rapidly delocalizes over the five carbon unit. Addition of dioxygen yields a peroxyl radical, which is finally reduced to the hydroperoxide product	Rippkaea orientalis PCC 8801	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
linoleate + O2	-	Rippkaea orientalis PCC 8801	(9Z,11R,12Z)-11-hydroperoxyoctadeca-9,12-dienoate	-	?
linoleate + O2	analysis of substrate conformation and environment, overview	Rippkaea orientalis PCC 8801	(9Z,11R,12Z)-11-hydroperoxyoctadeca-9,12-dienoate	-	?
additional information	CspLOX1 only produces conjugated hydroperoxyoctadeca-9,12-dienoate in a way that it imitates CspLOX2 and forms 11-hydroperoxyoctadeca-9,12-dienoate	Rippkaea orientalis PCC 8801	?	-	?

Synonyms

Synonyms	Comment	Organism
CspLOX2	-	Rippkaea orientalis PCC 8801
lipoxygenase 2	-	Rippkaea orientalis PCC 8801

Cofactor

Cofactor	Comment	Organism	Structure
iron cofactor	coordination of the iron cofactor, overview	Rippkaea orientalis PCC 8801

General Information

General Information	Comment	Organism
additional information	structure-activity analysis, overview. Crucial residues in the direct environment of the narrow active site that form a clamp-like structure include Tyr360, Gly401 Ile617, Ile379, Leu405, and Leu621. Molecular dynamics simulations support a major role of steric shielding of active site clamp. Structure-activity analysis, modeling, overview	Rippkaea orientalis PCC 8801