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Literature summary extracted from

  • Tucker, D.E.; Ghosh, M.; Ghomashchi, F.; Loper, R.; Suram, S.; St John, B.; Girotti, M.; Bollinger, J.G.; Gelb, M.H.; Leslie, C.C.
    Role of phosphorylation and basic residues in the catalytic domain of cytosolic phospholipase A2alpha in regulating interfacial kinetics and binding and cellular function (2009), J. Biol. Chem., 284, 9596-9611.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.1.52 phorbol myristate acetate PMA, 100 nM to cells induces translocation (maximum after 15 min) of enzyme to the Golgi apparatus and release of 4-5% of cell-associated arachidonic acid within 80 min, effect impaired by PKC inhibition and block of protein synthesis Mus musculus
3.1.1.52 serum 10% mouse serum to cells induces immediate translocation (within 60 seconds) of enzyme to the Golgi apparatus and release of 20% of cell-associated arachidonic acid within 80 min, effect depends on ERK and p38/MAPK pathway activation and calcium binding, effect is impaired by block of protein synthesis Mus musculus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.4 expression of full-length wild-type and mutant cPLA2alphas containing the affinity peptide YHHHHHH fused to the C-terminal Ala749, involving a recombinant adenovirus Homo sapiens
3.1.1.52 together with EGFP cDNA in AdEasy vector or together with monomeric EYFP or ECFP cDNA in pVQAd5CMVK-NpA shuttle plasmid for adenovirus-mediated infection of lung fibroblasts Mus musculus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.4 A206K site-directed mutagenesis Homo sapiens
3.1.1.4 D43N site-directed mutagenesis Homo sapiens
3.1.1.4 K488N/K543N/K544N site-directed mutagenesis, the mutant shows 2fold increased activity compared to the wild-type enzyme Homo sapiens
3.1.1.4 additional information reconstitution of immortalized mouse lung fibroblasts lacking endogenous cPLA2alpha, IMLF-/-, with human wild-type and cPLA2alpha mutants, overview Homo sapiens
3.1.1.4 S505A site-directed mutagenesis Homo sapiens
3.1.1.4 S505A/S727A site-directed mutagenesis Homo sapiens
3.1.1.4 S505A/S727A/S515A site-directed mutagenesis Homo sapiens
3.1.1.4 S515A site-directed mutagenesis Homo sapiens
3.1.1.4 S727A site-directed mutagenesis Homo sapiens
3.1.1.52 K271N/K273N/K274N wild-type activity, half the wild-type increase in activity in presence of phosphatidylinositol 4,5-biphosphate Mus musculus
3.1.1.52 K43N impaired release of arachidonic acid in response to serum and phorbol myristate acetate, no translocation to Golgi apparatus in response to serum, C2-domain mutant, calcium and membrane binding Mus musculus
3.1.1.52 K488N/K543N/K544N 2fold increased specific activity (independent of phosphatidylinositol 4,5-biphosphate), 78% reduced release of arachidonic acid and increased Golgi-binding in response to serum, PIP2-binding site triple mutant Mus musculus
3.1.1.52 K541N/K543N/K544N 2fold increased specific activity (independent of phosphatidylinositol 4,5-biphosphate) Mus musculus
3.1.1.52 S505A 38% reduced release of arachidonic acid and prolonged accumulation at Golgi apparatus in response to serum and 30% reduced release of arachidonic acid in response to phorbol myristate acetate, phosphorylation site mutant Mus musculus
3.1.1.52 S505A/S727A 30% reduced release of arachidonic acid in response to phorbol myristate acetate and 50% in response to serum which is further reduced by ERK inhibition, prolonged accumulation at Golgi apparatus in response to serum, phosphorylation site double mutant Mus musculus
3.1.1.52 S505A/S727A/S515A similar response to serum as double mutant S505A/S727A, phosphorylation site triple mutant Mus musculus
3.1.1.52 S515A phosphorylation site mutant Mus musculus
3.1.1.52 S727A similar release of arachidonic acid in response to serum like wild-type enzyme, phosphorylation site mutant Mus musculus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.1.4 membrane calcium, phosphorylation at Ser505 and the putative phosphatidylinositol 4,5-bisphosphate binding site regulate translocation and arachidonic acid release, overview Homo sapiens 16020
-
3.1.1.52 Golgi apparatus transient serum-induced translocation of EGFP-tagged enzyme (7%) in parallel to sharp increase in intracellular calcium (within initial 60 seconds), stable binding dependent on influx of extracellular calcium induced here by phorbol myristate acetate Mus musculus 5794
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.1.4 Ca2+ calcium, phosphorylation at Ser505 and the putative phosphatidylinositol 4,5-bisphosphate binding site regulate translocation and arachidonic acid release, overview. Influx of extracellular calcium is essential for stable binding of cPLA2alpha to Golgi and arachidonic acid release. phosphatidylinositol 4,5-bisphosphate improves Ca2+ binding by about 2fold Homo sapiens
3.1.1.52 Ca2+ 50% of maximal activation at 0.7 +/-0.1 microM (independent of presence of 10 mol% phosphatidylinositol 4,5-biphosphate), 12% less activity in absence of calcium Mus musculus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.1.4 additional information Homo sapiens cPLA2alpha is regulated by phosphorylation at Ser505 and calcium-induced translocation to membranes ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.4 Homo sapiens
-
-
-
3.1.1.52 Mus musculus
-
 wild-type or homozygous knockout mice
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.1.1.4 phosphoprotein calcium, phosphorylation at Ser505 and the putative phosphatidylinositol 4,5-bisphosphate binding site regulate translocation and arachidonic acid release, overview. Preparation of phosphorylated cPLA2alpha forms: cPLA2alpha-505P and cPLA2alpha-515P Homo sapiens
3.1.1.52 phosphoprotein phosphorylation at S505 increases activity 2fold, phosphorylation state not influenced by serum or phorbol myristate acetate, no impact on calcium-mediated membrane binding, constitutively phosphorylated at S505 in immortalized cells (SV40 transformed) Mus musculus

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.4 recombinant full-length wild-type and mutant cPLA2alphas containing the affinity peptide YHHHHHH Homo sapiens
3.1.1.52 following expression with C-terminal hexa-His-tag, dephosphorylation with potato acid phosphatase Mus musculus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.1.1.4 blood platelet
-
 Homo sapiens
-
3.1.1.52 blood platelet activated Mus musculus
-
3.1.1.52 lung fibroblast isolated from wild-type or knockout mice, subsequently SV40 transformed Mus musculus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.1.52 additional information
-
 5fold increase in specific activity of activated platelet-derived wild-type and phosphorylated forms in presence of 10 mol% phosphatidylinositol 4,5-biphosphate Mus musculus
3.1.1.52 0.21
-
 mutant K271N/K273N/K274N, from activated platelets Mus musculus
3.1.1.52 0.23
-
 wild-type, from activated platelets Mus musculus
3.1.1.52 0.26
-
 phoshorylated at residue S515, from activated platelets Mus musculus
3.1.1.52 0.38
-
 phoshorylated at residue S505, from activated platelets Mus musculus
3.1.1.52 0.39
-
 mutant K488N/K543N/K544N, from activated platelets, in presence of 10 mol% phosphatidylinositol 4,5-biphosphate Mus musculus
3.1.1.52 0.4
-
 mutant K488N/K543N/K544N, from activated platelets Mus musculus
3.1.1.52 0.405
-
 phosphorylated at residues S505 and S727, from activated platelets Mus musculus
3.1.1.52 0.68
-
 mutant K271N/K273N/K274N, from activated platelets, in presence of 10 mol% phosphatidylinositol 4,5-biphosphate Mus musculus
3.1.1.52 0.866
-
 phoshorylated at residue S515, from activated platelets, in presence of 10 mol% phosphatidylinositol 4,5-biphosphate Mus musculus
3.1.1.52 1.03
-
 wild-type, from activated platelets, in presence of 10 mol% phosphatidylinositol 4,5-biphosphate Mus musculus
3.1.1.52 1.45
-
 phoshorylated at residue S505, from activated platelets, in presence of 10 mol% phosphatidylinositol 4,5-biphosphate Mus musculus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.4 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) + H2O
-
 Homo sapiens 1-palmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) + palmitic acid
-
 ?
3.1.1.4 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphocholine + H2O
-
 Homo sapiens 1-palmitoyl-sn-glycero-3-phosphocholine + arachidonic acid
-
 ?
3.1.1.4 1-stearoyl-2-arachidonyl-phosphatidylinositol 4,5-bisphosphate + H2O
-
 Homo sapiens 1-stearoyl-sn-glycero-3-phosphoinositol 4,5-bisphosphate + arachidonic acid
-
 ?
3.1.1.4 additional information cPLA2alpha is regulated by phosphorylation at Ser505 and calcium-induced translocation to membranes Homo sapiens ?
-
 ?
3.1.1.4 additional information interfacial binding and kinetics of cPLA2 and its phosphorylated forms in vitro, lipid vesicle formation, overview Homo sapiens ?
-
 ?
3.1.1.52 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphocholine + H2O 2 min, 37°C (pre-warmed), in presence of 0.5 mg/ml bovine serum albumin and various concentrations of calcium Mus musculus arachidonic acid + 1-palmitoyl-sn-glycero-3-phosphocholine quantification of release of radio-labelled arachidonic acid ?

Synonyms

EC Number Synonyms Comment Organism
3.1.1.4 cPLA2alpha
-
 Homo sapiens
3.1.1.4 group IVA cytosolic phospholipase A2
-
 Homo sapiens
3.1.1.4 phospholipase A2alpha
-
 Homo sapiens
3.1.1.4 PLA2alpha
-
 Homo sapiens
3.1.1.52 cPLA2alpha
-
 Mus musculus
3.1.1.52 group IVA cytosolic phospholipase A2alpha
-
 Mus musculus