Literature summary for 1.13.11.31 extracted from

  • Aleem, A.M.; Jankun, J.; Dignam, J.D.; Walther, M.; Kuehn, H.; Svergun, D.I.; Skrzypczak-Jankun, E.
    Human platelet 12-lipoxygenase, new findings about its activity, membrane binding and low-resolution structure (2008), J. Mol. Biol., 376, 193-209.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged wild-type and mutant enzymes in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
low-resolution 16 A structure of enzyme in solution, dynamic light scattering and SAXS analysis of hp-12LOX shows that this enzyme as a whole is stable in the dimeric form, while mutants without PLAT domains associate into larger aggregates Homo sapiens

Engineering

Protein Variants Comment Organism
additional information truncation of the N-terminal PLAT domain impairs catalytic activity but does not alter reaction specificity Homo sapiens
C7S site-directed mutagenesis, the mutant shows highly reduced iron content compared to the wild-type enzyme, but exchange of the surface-exposed cysteine alters neither the catalytic activity nor the reaction specificity Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information dependence of the recombinant His-tagged isozyme hp-12LOX enzymatic reaction rate on the concentration of arachidonic acid and pH, kinetics, overview Homo sapiens
0.0072
-
arachidonic acid cytosolic hp-12LOX, pH 7.4, recombinant wild-type enzyme Homo sapiens
0.0087
-
arachidonic acid membrane hp-12LOX, pH 7.4, recombinant wild-type enzyme Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane membrane binding activities of wild-type platelet 12-lipoxygenase and mutant enzymes at pH 7.4, role of the PLAT domain membrane binding, overview Homo sapiens
-
-
cytosol platelet 12-lipoxygenase Homo sapiens
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ iron content of wild-type and mutant enzymes, overview Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
158000
-
about, recombinant His-tagged wild-type and mutant enzymes, gel filtration Homo sapiens
76500
-
2 * 76500, recombinant His-tagged enzyme, SDS-PAGE Homo sapiens
675000
-
about, recombinant no-PLAT-mutant, gel filtration Homo sapiens
62650
-
x * 62650, recombinant no-PLAT-mutant, SDS-PAGE Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
arachidonate + O2 Homo sapiens
-
(5Z,8Z,10E,14Z)-(12S)-12-hydroperoxyeicosa-5,8,10,14-tetraenoate further metabolized to 12(S)-hydroxyeicosatetraenoic acid ?

Organism

Organism UniProt Comment Textmining
Homo sapiens P18054
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
arachidonate + O2
-
Homo sapiens (5Z,8Z,10E,14Z)-(12S)-12-hydroperoxyeicosa-5,8,10,14-tetraenoate further metabolized to 12(S)-hydroxyeicosatetraenoic acid ?
arachidonate + O2 two molecules of arachidonic acid bind to one molecule of the enzyme, role of the PLAT domain in activity and membrane binding, overview Homo sapiens (5Z,8Z,10E,14Z)-(12S)-12-hydroperoxyeicosa-5,8,10,14-tetraenoate
-
?

Subunits

Subunits Comment Organism
dimer 2 * 76500, recombinant His-tagged enzyme, SDS-PAGE Homo sapiens
oligomer x * 62650, recombinant no-PLAT-mutant, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
hp-12LOX
-
Homo sapiens
human platelet 12-lipoxygenase
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Homo sapiens

pH Range

pH Minimum pH Maximum Comment Organism
7.4 8.7
-
Homo sapiens