Phospholipase d activity facilitates ca2+-induced aggregation and fusion of complex liposomes. Blackwood, R. Alexander, James E. Smolen, Amy Transue, Ronald J. Hessler, Donna M. Harsh, Ruth C. Brower, and Scott French. University of Michigan Medical Center, Ann Arbor, Michigan 48109 -0244, Departmaent of Pediatrics, Divisions of Infectious Diseases and Hematology
APStracts 3:0388C, 1996.
Phospholipase D activation in stimulated neutrophils results in the conversion of membrane phosphatidylcholine (PC) to phosphatidic acid (PA). This change in membrane phospholipid composition has two potentially positive effects on degranulation: 1) it replaces a non -fusogenic phospholipid with a fusogenic one; and 2) it increases the potential for interactions between membranes and the annexins. Modeling neutrophil degranulation, we examined the effect of phospholipase D (Streptomyces chromofuscus) hydrolysis on the aggregation and fusion of liposomes in the presence and absence of annexin I. We found that PLD-mediated conversion of PC to PA lowered the [Ca2+] required for fusion. Annexin I increased the rate of fusion in the presence of PA, although it did not lower threshold [Ca2+], which remained above the physiologic range. However, following hydrolysis by PLD, annexin I lowered the [Ca2+] required for aggregation by almost 3 orders of magnitude to near physiologic concentrations. These studies indicate that the activation of phospholipase D and the production of PA may play a role in annexin -mediated membrane-membrane apposition.

Received 5 August 1996; accepted in final form 21 October 1996.
APS Manuscript Number C439-6.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 31 December 1996