Cloning and Expression of Cytosolic Phospholipase A2 (cPLA2) and a Naturally Occurring Variant
作者： Kenneth R. Bundell , Lynne Douglas , John Morten , Maurice Needham , Robert D. Gordon
DOI: 10.1111/J.1432-1033.1996.0690W.X
关键词: Mitogen-activated protein kinase kinase 、 Biology 、 Cyclin-dependent kinase 2 、 Phosphorylation 、 Protein kinase A 、 MAP2K7 、 Biochemistry 、 MAP kinase kinase kinase 、 Kinase 、 Dephosphorylation 、 Molecular biology
摘要: Full-length cytosolic phopholipase A2 (cPLA2) was cloned from U937 cells and polymorphonuclear leukocytes (PMNLs) while a naturally occurring variant of cPLA2 which lacks residues Val473–Ala749 but has C-terminal extension ILMNLSEYMLWMSKVKRFM (DcPLA2) PMNLs mononuclear leukocytes. We were unable to clone DcPLA2 cells. When expressed in insect cells, both proteins detected cell lysates by SDS/PAGE as single bands apparent molecular masses 100 kDa 57 kDa, respectively. active lysophospholipase assays inactive assays. phosphorylated stoichiometrically p42 mitogen–activated protein (MAP) kinase vitro at similar rate other physiological substrates this the major site phosphorylation identified amino acid sequencing Ser505. [32P]Ser(P)505 only dephosphorylated slow mammalian tissue homogenates. Protein phosphatases 2A, 2B 2C all contributed significantly overall dephosphorylation cPLA2. The MAP correlated with an approximately 1.5-fold increase specific enzyme activity reversed dephosphorylation.
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