Is phospholipase D really an enzyme? A comparison of in situ and in vitro activities.
Leaf phospholipase D activity was compared in vitro and in situ. In the in situ reaction stimulated by methanol only phosphatidylcholine and phosphatidylethanolamine were degraded until approx. 80% of these endogenous substrates had been consumed. Only then was a limited amount (approx. 20%) of endogenous phosphatidylglycerol degraded. Endogenous phosphatidylinositol was apparently not susceptible to phospholipase D in situ. In the vitro reaction the relative susceptibilities to degradation of added phospholipid substrates were (a) in the absence of "activators" phosphatidylethanolamine greater than phosphatidylglycerol greater than phosphatidylcholine, (b) in the presence of diethyl ether phosphatidylcholine greater than phosphatidylethanolamine greater than phosphatidylglycerol and (c) in the presence of sodium dodecyl sulphate phosphatidylcholine greater than phosphatidylethanolamine = phosphatidylglycerol. Minimum rates calculated for the in situ reaction in cauliflower leaf were 5-fold higher than maximum in vitro rates reported for the same material. Phospholipase D activity has been demonstrated by the in situ reaction in all leaf tissue so far examined. From these data we conclude that phospholipase D may be an integral part of membranes containing phosphatidylcholine and phosphatidylethanolamine, but not of membranes containing phosphatidylglycerol. We also suggest that phospholipase D may not be a physiological enzyme, but rather a structural protein of phosphatidylcholine- and phosphatidylethanolamine-containing membranes and which, under certain non-physiological conditions, possess enzymic properties.
D002725:Chloroform / Q000494:pharmacology; D002736:Chloroplasts / Q000187:drug effects / Q000378:metabolism; D004986:Ether / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism; D066298:In Vitro Techniques; D000432:Methanol / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D010740:Phospholipases / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D010944:Plants / Q000187:drug effects / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D005227:Fatty Acids; D010743:Phospholipids; D004986:Ether; D012967:Sodium Dodecyl Sulfate; D002725:Chloroform; D010740:Phospholipases; D000432:Methanol