Cardiolipin hydrolysis by human phospholipases A2. The multiple enzymatic activities of human cytosolic phospholipase A2.
The ability of mammalian phospholipases A2 (PLA2) to hydrolyse cardiolipin (diphosphatidylglycerol) was monitored with a fluorescent displacement assay which allows the use of natural phospholipid substrates. The mammalian enzymes used were porcine pancreatic (Group I) secretory PLA2 (sPLA2), human non-pancreatic (Group II) sPLA2 and human cytosolic PLA2 (cPLA2). High activity was observed with porcine pancreas sPLA2 whereas the human sPLA2 demonstrated only minimal activity with this substrate. In comparison, sPLA2 from Naja naja venom (Group I) also showed only modest activity with this substrate. Since many lipases possess PLA1 activity, a representative enzyme from Rhizopus arrhizus was also assessed for its ability to hydrolyse cardiolipin which proved to be a good substrate for this fungal lipase. In all cases dilysocardiolipin was the major product while some monolyso intermediate was detected after chromatographic separation. Human cPLA2 was unable to hydrolyse cardiolipin at a significant rate, however, both monolysocardiolipin and dilysocardiolipin, which are prepared by the PLA2-catalysed hydrolysis of cardiolipin, were good substrates providing a further example of the extensive lysophospholipase activity of this enzyme. Moreover, cardiolipin that was initially hydrolysed in situ with either excess porcine pancreatic PLA2 or R. arrhizus lipase (PLA1) was subsequently hydrolysed by human cPLA2. One explanation of this result is that human cPLA2 is able to hydrolyse both 1-acyl and 2-acyl-lysophospholipids. (c) 1998 Elsevier Science B.V.
Buckland
A G
AG
Department of Biochemistry, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK.
Kinkaid
A R
AR
Wilton
D C
DC
D002308:Cardiolipins / Q000378:metabolism*; D002460:Cell Line; D002855:Chromatography, Thin Layer; D003600:Cytosol / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D008049:Lipase / Q000378:metabolism; D008246:Lysophospholipids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054466:Phospholipases A1; D054467:Phospholipases A2
D002308:Cardiolipins; D008246:Lysophospholipids; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; C067187:dilysocardiolipin; C067188:monolysocardiolipin; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol; D008049:Lipase; D010741:Phospholipases A; D054466:Phospholipases A1; D054467:Phospholipases A2; C017251:1,2-oleoylphosphatidylcholine