Abstract
The phospholipid composition was determined for the amebocyte of the primitive arthropod Limulus polyphemus. The total fatty acid composition of the cells' lipids was analyzed by gas chromatography/mass spectrometry (GC/MS) of fatty acid methyl esters (FAME). The FAME analysis revealed high levels of 20-carbon polyunsaturated fatty acids (PUFA), especially arachidonic (20∶4n-6) and eicosapentaenoic (20∶5n-3) acids. Almost 20% of the total lipid profile was comprised of dimethyl acetals of 16- to 20-carbon chain lengths, indicative of plasmalogens in the phospholipid pool. Phospholipids, analyzed by high-pressure liquid chromatography, included phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylinositol (PI), sphingomyelin (SPH), and cardiolipin (CL). PE and PC levels predominated at 42.2 and 36.3%, respectively. Smaller amounts of PS (9.0%) and PI (6.2%) were present, as well as low levels of SPH (4.6%), CL (1.6%), and trace amounts of lysophosphatidylcholine. The major phospholipid species, PE, PC, PS and PI, were collected and their molecular species were examined by electrospray-ionization mass spectrometry. The molecular species within the phospholipid classes reflected the high levels of PUFA seen in the total lipid profile. PI was mainly composed of 18∶0a/20∶4. Over half of the PS consisted of 18∶0a/18∶1 and 18∶0a/20∶4. The major PE species were 20∶1p/20∶5, 20∶1p/20∶4, 18∶0p/20∶5, and 18∶0p/20∶4. PC had the largest distribution of molecular species, and its most abundant species were 16∶0e/20∶5, 16∶0e/20∶4, and 16∶0p/20∶4. The presence of 16∶0e/20∶4 is the first documentation of a specific precursor to platelet-activating factor in an invertebrate hemocyte. Note: at the sn-1 position: [a=1=O-acyl, e=1-O-alkylether, and p=1-O-alk-1′-enyl (plasmalogen)].
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Abbreviations
- AA:
-
arachidonic acid
- CL:
-
cardiolipin
- DMA:
-
dimethyl acetal(s)
- EPA:
-
eicosapentaenoic acid
- ESI/MS:
-
electrospray-ionization/mass spectrometry
- FAME:
-
fatty acid methyl ester(s)
- GC/MS:
-
gas chromatography/mass spectrometry
- HPLC:
-
high-pressure liquid chromatography
- IP3 :
-
inositol-1,4,5-triphosphate
- LPC:
-
lysophosphatidylcholine
- PAF:
-
platelet-activating factor
- PC:
-
phosphatidylcholine
- PE:
-
phosphatidylethanolamine
- PI:
-
phosphatidylinositol
- PS:
-
phosphatidylserine
- PUFA:
-
polyunsaturated fatty acid(s)
- SPH:
-
sphingomyelin; phospholipid molecular species nomenclature, from Reference, 1, n:jk/s:t, where n=number of carbons at the sn-1 position, j=number of double bonds in the sn-1 chain, k=the type of linkage at the sn-1 position [a=1-O-acyl, e=1-O-alkylether, and p=1-O-alk-1′-enyl (plasmalogen)], s=number of carbons at the sn-2 position, and t=the number of double bonds in the sn-2 chain
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MacPherson, J.C., Pavlovich, J.G. & Jacobs, R.S. Phospholipid composition of the granular amebocyte from the horseshoe crab, Limulus polyphemus . Lipids 33, 931–940 (1998). https://doi.org/10.1007/s11745-998-0290-y
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DOI: https://doi.org/10.1007/s11745-998-0290-y