Literature summary for 3.1.1.52 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.

Search for more literature for 3.1.1.52

Activating Compound

Activating Compound	Comment	Organism	Structure
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
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)

Cloned (Comment)	Organism
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

Protein Variants	Comment	Organism
K271N/K273N/K274N	wild-type activity, half the wild-type increase in activity in presence of phosphatidylinositol 4,5-biphosphate	Mus musculus
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
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
K541N/K543N/K544N	2fold increased specific activity (independent of phosphatidylinositol 4,5-biphosphate)	Mus musculus
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
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
S505A/S727A/S515A	similar response to serum as double mutant S505A/S727A, phosphorylation site triple mutant	Mus musculus
S515A	phosphorylation site mutant	Mus musculus
S727A	similar release of arachidonic acid in response to serum like wild-type enzyme, phosphorylation site mutant	Mus musculus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
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

Metals/Ions	Comment	Organism	Structure
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

Organism

Organism	UniProt	Comment	Textmining
Mus musculus	-	wild-type or homozygous knockout mice	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
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)

Purification (Comment)	Organism
following expression with C-terminal hexa-His-tag, dephosphorylation with potato acid phosphatase	Mus musculus

Source Tissue

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
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

Synonyms	Comment	Organism
cPLA2alpha	-	Mus musculus
group IVA cytosolic phospholipase A2alpha	-	Mus musculus

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Release 2023.1 (January 2023)