Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.

The synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine, 1-O-[12-(2-naphthyl)-dodec-11-enyl]-2-O-decanoyl-sn-glycerol-3- phosphocholine (NVPC), is described. This involves a Wittig reaction between 2-naphthaldehyde and a phosphonium salt which gives the trans-naphthylvi...

Full description

Bibliographic Details
Main Authors: H S Hendrickson, E K Hendrickson, T J Rustad
Format: Article
Language:English
Published: Elsevier 1987-07-01
Series:Journal of Lipid Research
Online Access:http://www.sciencedirect.com/science/article/pii/S0022227520386533
id doaj-5bb30b6893464b9ba731ada93af18259
record_format Article
spelling doaj-5bb30b6893464b9ba731ada93af182592021-04-25T04:20:22ZengElsevierJournal of Lipid Research0022-22751987-07-01287864872Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.H S HendricksonE K HendricksonT J RustadThe synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine, 1-O-[12-(2-naphthyl)-dodec-11-enyl]-2-O-decanoyl-sn-glycerol-3- phosphocholine (NVPC), is described. This involves a Wittig reaction between 2-naphthaldehyde and a phosphonium salt which gives the trans-naphthylvinyl group as the predominant isomer. Lyso NVPC was prepared from NVPC by phospholipase A2 action. NVPC absorbs strongly at 248 nm (epsilon = 58,300 M-1 cm-1) and gives broad fluorescence emission with maxima at 343 nm and 360 nm and a quantum yield of 0.10 in ethanol. An assay for phospholipase A2 was developed using high performance liquid chromatography with fluorescence detection to separate and quantify NVPC and lyso NVPC. Activities as low as 1-2 pmol/min in an assay volume of 0.1 ml can easily be measured. The assay was used with a pure enzyme from cobra venom and a crude enzyme from synovial fluid. Enzyme specificities for phosphatidylcholine and NVPC with cobra venom and porcine pancreatic phospholipases A2 were compared using a titrametric assay. The use of the assay with NVPC to study the metabolism of platelet activating factor is discussed.http://www.sciencedirect.com/science/article/pii/S0022227520386533
collection DOAJ
language English
format Article
sources DOAJ
author H S Hendrickson
E K Hendrickson
T J Rustad
spellingShingle H S Hendrickson
E K Hendrickson
T J Rustad
Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.
Journal of Lipid Research
author_facet H S Hendrickson
E K Hendrickson
T J Rustad
author_sort H S Hendrickson
title Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.
title_short Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.
title_full Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.
title_fullStr Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.
title_full_unstemmed Synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase A2.
title_sort synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine and its use in the assay of phospholipase a2.
publisher Elsevier
series Journal of Lipid Research
issn 0022-2275
publishDate 1987-07-01
description The synthesis of a naphthylvinyl-labeled glycerol ether analog of phosphatidylcholine, 1-O-[12-(2-naphthyl)-dodec-11-enyl]-2-O-decanoyl-sn-glycerol-3- phosphocholine (NVPC), is described. This involves a Wittig reaction between 2-naphthaldehyde and a phosphonium salt which gives the trans-naphthylvinyl group as the predominant isomer. Lyso NVPC was prepared from NVPC by phospholipase A2 action. NVPC absorbs strongly at 248 nm (epsilon = 58,300 M-1 cm-1) and gives broad fluorescence emission with maxima at 343 nm and 360 nm and a quantum yield of 0.10 in ethanol. An assay for phospholipase A2 was developed using high performance liquid chromatography with fluorescence detection to separate and quantify NVPC and lyso NVPC. Activities as low as 1-2 pmol/min in an assay volume of 0.1 ml can easily be measured. The assay was used with a pure enzyme from cobra venom and a crude enzyme from synovial fluid. Enzyme specificities for phosphatidylcholine and NVPC with cobra venom and porcine pancreatic phospholipases A2 were compared using a titrametric assay. The use of the assay with NVPC to study the metabolism of platelet activating factor is discussed.
url http://www.sciencedirect.com/science/article/pii/S0022227520386533
work_keys_str_mv AT hshendrickson synthesisofanaphthylvinyllabeledglyceroletheranalogofphosphatidylcholineanditsuseintheassayofphospholipasea2
AT ekhendrickson synthesisofanaphthylvinyllabeledglyceroletheranalogofphosphatidylcholineanditsuseintheassayofphospholipasea2
AT tjrustad synthesisofanaphthylvinyllabeledglyceroletheranalogofphosphatidylcholineanditsuseintheassayofphospholipasea2
_version_ 1721510403945529344