| Synthesis and applications of stereospecifically (3)H-labeled arachidonic acids as mechanistic probes for lipoxygenase and cyclooxygenase catalysis. | |
| | |
MedLine Citation:
|
PMID: 10933865 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Stereospecifically (3)H-labeled substrates are useful tools in studying the mechanism of hydrogen abstractions involved in the oxygenation of polyunsaturated fatty acids. Here, we describe modified methods for the synthesis of arachidonic acids labeled with a single chiral tritium on the methylene groups at carbons 10 or 13. The appropriate starting material is a ketooctadecanoic acid which is prepared from an unsaturated C18 fatty acid precursor or by total synthesis. The (3)H label is introduced by NaB(3)H(4) reduction and the resulting tritiated hydroxy fatty acid then is tosylated, separated into the enantiomers by chiral phase HPLC, and subsequently transformed into stearic acids. A variety of stereospecifically labeled unsaturated fatty acids are obtained using literature methods of microbial transformation with the fungus Saprolegnia parasitica. Two applications are described: (i) In incubations of [10S-(3)H]- and [10R-(3)H]arachidonic acids in human psoriatic scales we show that a 12R-lipoxygenase accounts not only for synthesis of the major product 12R-HETE, but it contributes also, through subsequent isomerization, to the minor amounts of 12S-HETE. (ii) The [10R-(3)H]- and [10S-(3)H]arachidonic acids were also used to demonstrate that prostaglandin ring formation by cyclooxygenases does not involve carbocation formation at C-10 of arachidonic acid as was hypothesized recently. |
| | |
Authors:
|
C Schneider; W E Boeglin; S Lai; J K Cha; A R Brash |
Related Documents
:
|
1858085 - Effect of inhaled 15-(s)-hydroxyeicosatetraenoic acid on tracheobronchial clearance in ... 15145985 - Human cyp4f3s are the main catalysts in the oxidation of fatty acid epoxides. 7793625 - Analysis of leukotrienes, lipoxins, and monooxygenated metabolites of arachidonic acid ... 20732035 - Microcystin-induced activation of prostaglandin synthesis and phospholipid metabolism i... 23096425 - Signal transduction in leaf senescence. 18590915 - Cyclic gmp signaling in cardiomyocytes modulates fatty acid trafficking and prevents tr... |
Publication Detail:
|
Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
|
Title: Analytical biochemistry Volume: 284 ISSN: 0003-2697 ISO Abbreviation: Anal. Biochem. Publication Date: 2000 Aug |
Date Detail:
|
Created Date: 2000-10-02 Completed Date: 2000-10-02 Revised Date: 2007-11-14 |
Medline Journal Info:
|
Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 125-35 Citation Subset: IM |
Copyright Information:
|
Copyright 2000 Academic Press. |
Affiliation:
|
Division of Clinical Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6602, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Arachidonic Acids / chemical synthesis, chemistry* Carbon / metabolism Chromatography, High Pressure Liquid Cyclooxygenase 2 Dinoprostone / chemistry Humans Hydrogen / metabolism Hydroxyeicosatetraenoic Acids / biosynthesis Isoenzymes / metabolism Lipoxygenase / metabolism* Membrane Proteins Mice Models, Chemical Prostaglandin-Endoperoxide Synthases / metabolism* Psoriasis / metabolism Stereoisomerism Time Factors |
| Grant Support | |
ID/Acronym/Agency:
|
GM-53638/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Arachidonic Acids; 0/Hydroxyeicosatetraenoic Acids; 0/Isoenzymes; 0/Membrane Proteins; 1333-74-0/Hydrogen; 363-24-6/Dinoprostone; 7440-44-0/Carbon; EC 1.13.11.12/Lipoxygenase; EC 1.14.99.1/Cyclooxygenase 2; EC 1.14.99.1/PTGS2 protein, human; EC 1.14.99.1/Prostaglandin-Endoperoxide Synthases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Quantification of active caspase 3 in apoptotic cells.
Next Document: High-performance cation-exchange chromatography and pulsed amperometric detection for the separation...