Cytosolic phospholipase A2 from U937 cells: size of the functional enzyme by radiation inactivation.
We have studied the cytosolic phospholipase A2 (cPLA2) of human U937 cells by radiation inactivation in order to characterize the functional form of the native enzyme by a method that was independent of the discrepancies observed by SDS-PAGE and cDNA cloning. The Radiation Inactivation Size of cPLA2 was reproducible and gave a value of 76,800-80,100 daltons. We eluted the active enzyme from polyacrylamide-gradient gel electrophoresis at a molecular weight of 77,000, confirming the irradiation result. We conclude that cPLA2 is active as the monomeric enzyme and is composed of a single major functional domain that is sensitive to irradiation.
Tremblay
N M
NM
Merck Frosst Centre for Therapeutic Research, Merck Frosst Canada Inc., Dorval, Québec.
Nicholson
D
D
Potier
M
M
Weech
P K
PK
D002478:Cells, Cultured / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008970:Molecular Weight; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification; D054467:Phospholipases A2; D011487:Protein Conformation; D011839:Radiation, Ionizing
D010741:Phospholipases A; D054467:Phospholipases A2