Mechanism of regulation of group IVA phospholipase A2 activity by Ser727 phosphorylation.
Although group IVA cytosolic phospholipase A(2) (cPLA(2)alpha) has been reported to be phosphorylated at multiple Ser residues, the mechanisms by which phosphorylation at different sites regulates cPLA(2)alpha activities are not fully understood. To explore the possibility that phosphorylation of Ser(727) modulates cellular protein-protein interactions, we measured the effect of Ser(727) mutations on the interaction of cPLA(2)alpha with a reported cPLA(2)alpha-binding protein, p11. In vitro activity assays and membrane binding measurements by surface plasmon resonance analysis showed that a heterotetramer (A2t) of p11 and annexin A2, but not p11 or annexin A2 alone, directly binds cPLA(2)alpha via Ser(727), which keeps the enzyme from binding the membrane and catalyzing the phospholipid hydrolysis. Phosphorylation of Ser(727) disrupts this inhibitory cPLA(2)alpha-A2t interaction, thereby activating cPLA(2)alpha. Subcellular translocation and activity measurements in HEK293 cells cotransfected with cPLA(2)alpha and p11 also showed that p11, in the form of A2t, inhibits cPLA(2)alpha by the same mechanism and that phosphorylation of Ser(727) activates cPLA(2)alpha by interfering with the inhibitory cPLA(2)alpha-A2t interaction. Collectively, these studies provide new insight into the regulatory mechanism of cPLA(2)alpha through Ser(727) phosphorylation.
Tian
Wen
W
Department of Chemistry, University of Illinois, Chicago, Illinois 60607, USA.
Wijewickrama
Gihani T
GT
Kim
Jung Hwan
JH
Das
Sudipto
S
Tun
Moe P
MP
Gokhale
Nikhil
N
Jung
Jin Woo
JW
Kim
Kwang Pyo
KP
Cho
Wonhwa
W
D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D054467:Phospholipases A2 / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism*; D018411:Spodoptera; D020349:Surface Plasmon Resonance; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D012694:Serine; D015759:Ionomycin; D054467:Phospholipases A2; D013755:Tetradecanoylphorbol Acetate