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Sterol and steryl ester regulation of phospholipase A2 from the mosquito parasiteLagenidium giganteum

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Lipids

Abstract

Lagenidium giganteum, a facultative parasite of mosquito larvae, cannot synthesize sterols, and requires an exogenous source of these lipids in order to enter its reproductive cycle. This parasite grows vegetatively in the absence of sterols, but requires cholesterol or structurally related compounds to produce motile zoospores, which are the only stage capable of infecting mosquitoes. Sterols structurally related to cholesterol and some steryl esters inhibited the activity ofL. giganteum phospholipase A2 (PLA2), an enzyme that hydrolyzes fatty acids from thesn-2 position of glycerophospholipids. Sterols that induce reproduction inhibitedL. giganteum PLA2 activity, while sterols and steroids that do not support sporulation had minimal effect. Most steryl esters had no effect on enzyme activity, but cholesteryl arachidonate (CA) was a potent inhibitor of parasite PLA2. Not all enzymes partly purified using a DEAE-Sephacel column were affected by these lipids, demonstrating selective inhibition of specific enzymes. Potency was enhanced by up to several orders of magnitude if epoxy fatty acids were esterified to the cholesterol nucleus. The steryl ester pool was dynamic during morphogenesis, and the fatty acid composition of the steryl esters did not mimic total cell or membrane (glycerophospholipid) fatty acid composition asL. giganteum proceeded through its growth cycle. Synthesis of CA and monoepoxy CA by the parasite was confirmed using electrospray mass spectrometry and collision-induced dissociation. Steryl derivatives selectively inhibited PLA2 enzymes from bovine pancreas, snake venom, and human cytoplasmic 85-kDa PLA2.

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Abbreviations

CAE:

cholesteryl arachidonate epoxides

ESI-MS:

electrospray-mass spectrometry

ESI-MS/MS:

electrospray tandem mass spectrometry (collision-induced dissociation)

FAME:

fatty acid methyl esters

GC:

gas chromatography

GPL:

glycerophospholipids

PLA2 :

phospholipase A2

cholesteryl-9-HODE:

cholesteryl-9-hydroxy-10,12-octadecadienoic acid

cholesteryl-13-HODE:

cholesteryl-13-hydroxy-9,11-octadecadienoic acid

SE:

steryl esters

TLC:

thin-layer chromatography

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Author notes

  1. Joanna K. MacKichan

    Present address: Department of Microbiology and Immunology, Stanford University, Stanford, CA

Authors and Affiliations

  1. Botany Department 351330, University of Washington, 98195-1330, Seattle, Washington

    James L. Kerwin, Joanna K. MacKichan, Melinda J. Semon, Alison M. Wiens, Carolyn C. DeRose & Jessica J. Torvik

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Kerwin, J.L., MacKichan, J.K., Semon, M.J. et al. Sterol and steryl ester regulation of phospholipase A2 from the mosquito parasiteLagenidium giganteum . Lipids 31, 1179–1188 (1996). https://doi.org/10.1007/BF02524293

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