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Localization of long-chain fatty acids and unconventional sterols in spherulous cells of a marine sponge

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Lipids

Abstract

The first direct evidence is provided for the presence of unconventional lipids in a particular subcellular membrane system of a sponge. Spherulous cells were isolated from the variety of cell types present in the marine spongeAplysina fistularis by density gradient centrifugation. Spherulous cell plasma membrane was subsequently isolated by cell rupture followed by differential centrifugation and sucrose, or Percoll, density gradient ultracentrifugation. Plasma membrane isolates were identified and assessed for purity using [3H] concanavalin A plasma membrane marker, sodium dodecyl sulfate polyacrylamide gel electrophoresis and ratios of protein, sterol and phosphate. Plasma membrane isolates could not be assessed for purity by traditional enzymatic means. Spherulous cell plasma membrane was found to contain unusual lipids, including long-chain (C24–C30) fatty acids (16.8–27.2%) and unconventional 26-alkylated sterols (66.4–72.6%), in addition to more conventional fatty acids and sterols. Spherulous cell intracellular membranes were also found to contain long-chain fatty acids and unconventional sterols, although the relative importance of these unusual lipids apparently varies between intracellular membranes, with some containing approximately 50% long-chain acids.

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Abbreviations

BSA:

bovine serum albumin

CMF-ASW:

calcium-and magnesium-free artificial sea water

[3H]con A:

[3H]concanavalin A

EGTA:

ethylene glycol bis(β-aminoethyl ether)N,N′-tetraacetic acid

HM:

homogenizing medium

LB:

labeling buffer

LCFA:

long-chain fatty acid

P1, P2 and P3:

pellets obtained by respective centrifuging at 1075, 15,000 and 100,000×g

PUFA:

polyunsaturated fatty acid

SCFA:

short-chain fatty acid

SDS:

sodium dodecyl sulfate

ECL:

equivalent chain length

GC:

gas chromatography

HPLC:

high performance liquid chromatography

PAGE:

polyacrylamide gel electrophoresis

MS:

mass spectrometry

TLC:

thin layer chromatography

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Author information

Authors and Affiliations

  1. Department of Chemistry, Stanford University, 94305, Stanford, CA

    Mishelle P. Lawson, Janice E. Thompson & Carl Djerassi

Authors
  1. Mishelle P. Lawson
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  2. Janice E. Thompson
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  3. Carl Djerassi
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Additional information

Part 19 of “Phospholipid Studies of Marine Organisms.” For Part 18 see Li, H., Duzgunes, N., Ayanoglu, E., and Djerassi, C.,Chem. Phys. Lipids, in press.

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Lawson, M.P., Thompson, J.E. & Djerassi, C. Localization of long-chain fatty acids and unconventional sterols in spherulous cells of a marine sponge. Lipids 23, 1037–1048 (1988). https://doi.org/10.1007/BF02535649

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  • DOI: https://doi.org/10.1007/BF02535649

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