Abstract
Using chiral phase high-performance liquid chromatography of diacylglycerols, we have redetermined the ratios of 1,2-/2,3-diacyl-sn-glycerols resulting from acylation of 2-monoacylglycerols by membrane bound and solubilized triacylglycerol systhetase of rat intestinal mucosa. With 2-oleoyl[-3H]glycerol as the acyl acceptor and oleoyl-CoA as the acyl donor, 97–98% of the diacylglycerol product was 1,2(2,3)-dioleoyl-sn-glycerol, 90% of which was thesn-1,2-and 10% thesn-2,3-enantiomer. The remaining diacylglycerol (less than 3%) was thesn-1,3-isomer. The overall yield of acylation products was 70%, of which 60% were diacylglycerols and 40% triacylglycerols. With 2-oleylglycerol ether as the acyl acceptor and [1-14C]oleoyl-CoA as the acyl donor, 90% of the diradylglycerol was 1-oleoyl-2-oleyl-sn-glycerol and 10% was the 2-oleyl-3-oleoyl-sn-glycerol. The diradylglycerols made up 96% and the triradylglycerols 4% of the radioactive product. With 1-palmitoyl-sn-glycerol as the acyl acceptor and [1-14C]oleoyl-CoA as the acyl donor, the predominant reaction product was 1-palmitoyl-3-oleoyl-sn-glycerol. The 3-palmitoyl-sn-glycerol was not a suitable acyl acceptor. Both 1,2- and 2,3-diacyl-sn-glycerols were substrates for diacylglycerol acyltransferase as neither isomer was favored when 1,2-dioleoyl-rac-[2-3H]glycerol was used as the acyl acceptor. There was a marked decrease in the acylation of the 1(3)-oleoyl-2-oleyl-sn-glycerol to the 1,3-dioleoyl-2-oleyl-sn-glycerol. It is concluded that neither monoacylglycerol nor diacylglycerol acyltransferase exhibit absolute stereospecificity for acylglycerols as fatty acid acceptors.
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Abbreviations
- BCA:
-
bicinchonic acid
- CHAPS:
-
3-[(3-cholamidopropyl)dimethylammonio]propanesulfonate
- CI:
-
chemical ionization
- DG:
-
diradylglycerol
- DGAT:
-
diacylglycerol acyltransferase
- DNPU:
-
dinitrophenylurethane
- DTT:
-
dithiothreitol
- EDTA:
-
ethylenediaminetetraacetate (sodium salt)
- GLC:
-
gas-liquid chromatography
- HPLC:
-
high-performance liquid chromatography
- LC/MS:
-
liquid chromatography/mass spectrometry
- MG:
-
monoradylglycerol
- MGAT:
-
monoacylglycerol acyltransferase
- PMSF:
-
phenylmethylsulfonyl fluoride
- TG:
-
triradylglycerol
- TLC:
-
thin layer chromatography
- Tris:
-
tri[hydroxymethyl]aminomethane
- UV:
-
ultraviolet
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Lehner, R., Kuksis, A. & Itabashi, Y. Stereospecificity of monoacylglycerol and diacylglycerol acyltransferases from rat intestine as determined by chiral phase high-performance liquid chromatography. Lipids 28, 29–34 (1993). https://doi.org/10.1007/BF02536356
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DOI: https://doi.org/10.1007/BF02536356