Analysis of the stereochemistry of lipoxygenase-derived hydroxypolyenoic fatty acids by means of chiral phase high-pressure liquid chromatography

Kühn, Hartmut; Wiesner, Rainer; Ланкин, В. З.; Nekrasov, A. S.; Alder, Lutz; Schewe, Tankred

doi:10.1016/0003-2697(87)90609-9

Cited by 123 publications

(47 citation statements)

References 16 publications

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“…The assignment of the absolute configuration was based mM substrate in 100 mM borate buffer (pH 9.0 for sLOX and pH 7.0 on CD-spectroscopy in combination with chiral separa- (bLOX), and 5-HPANA (bLOX) showed a positive Cotton effect, so they have predominantly S-configuration (12). On this basis chiral separations were used to obtain the anandamide metabolites were eluted with methanol, dried under a accurate R/S ratios of the anandamide metabolites (Table nitrogen stream, redissolved in 1 ml hexane and hydrogenated with 3).…”

Section: Cyclooxygenase (6) Lipoxygenases (Linoleate: Oxygenmentioning

confidence: 99%

With Anandamide as Substrate Plant 5-Lipoxygenases Behave like 11-Lipoxygenases

Zadelhoff

Veldink

Vliegenthart

1998

Biochemical and Biophysical Research Communications

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stereo-selective dioxygenation of polyunsaturated fatty Anandamide, an endogenous ligand for cannabinoid acids with a (Z,Z)-1,4-pentadiene system. Porcine leureceptors CB1 and CB2, was incubated with purified kocyte 5-lipoxygenase was found inactive towards an-5-lipoxygenases from barley and tomato. This yielded andamide (4). In this study the potency of non-mamma-11S-hydroperoxy-5,8,12,14-eicosatetraenoylethanol-lian 5-lipoxygenases, like those from barley grains and amide (11S-HPANA) as major product (about 70%). tomato fruits, to use anandamide as substrate was inThis is in contrast with the dioxygenation of arachi-vestigated. Enzyme and substrate preparations. Lipoxygenases from barley grains (Triumph), tomato fruits (Trust) and soybeans (white Hilum) were purified as described (7-9). Anandamide and arachidonic acid were purified via solid phase extraction as follows: anandamide or In the early nineties anandamide (arachidonylethaarachidonic acid (5 mg) was dissolved in 2 ml methanol, and 50 ml nolamide) was isolated from porcine brain lipids and sodium borate buffer (pH 9.0) was added under continuous stirring.found to act as a ligand for cannabinoid receptors CB1 After lowering the pH to 4, the solution was applied to a SPE and CB2 (1,2). Although structurally distinct from can-(Bakerbond, 500 mg, J. T. Baker) column. Autooxidation products nabinoids, it shows a similar receptor binding affinity. were eluted with 5 ml methanol/water (80/20 v/v). Anandamide or arachidonic acid was eluted with 5 ml methanol, concentrated to 2Anandamide is an arachidonic acid derivative and ml under a stream of nitrogen and BHT was added to a final concenthereby a substrate for lipoxygenase (3-5) and for tration of 40 mM. cyclooxygenase (6). Lipoxygenases (linoleate: oxygenMetabolite generation. Each lipoxygenase was incubated with anoxidoreductase 1.13.11.12) are a group of non-heme andamide or arachidonic acid (final concentration 40 mM; 1 U lipoxyiron-containing enzymes which catalyze the regio-and genase per 3 mmol substrate) in 100 mM sodium borate buffer (pH 9.0 for the 15-lipoxygenase and pH 7.0 for the 5-lipoxygenases). The reaction was followed spectrophotometrically at 236 nm. After com-1 To whom correspondence should be addressed. Fax: (31) 30 pletion, the pH was lowered to 4 and the products were purified with 2540980. E-mail: veldink@cc.ruu.nl. SPE (Bakerbond, 500 mg, J. T. Baker) and then analyzed by HPLC Abbreviations used: sLOX, soybean lipoxygenase-1; bLOX, barley and/or GC/MS. lipoxygenase-1; tLOX, tomato lipoxygenase; SPE, solid phase extraction; RP-HPLC, reversed-phase high performance liquid chromatogSpectrophotometric assay. Spectrophotometric assays were performed by continuously monitoring the change in absorbance at 236 raphy; CP-HPLC, chiral-phase high performance liquid chromatography; GC/MS, gas chromatography/mass spectrometry; H(P)ANA, nm on a Hewlett-Packard 8452A diode array spectrophotometer. A molar absorbance of 23000 M 01 cm 01 at 236 nm was used for N-(2-hydroxyethyl)hydro(pero)xyarachidon...

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Section: Cyclooxygenase (6) Lipoxygenases (Linoleate: Oxygenmentioning

confidence: 99%

With Anandamide as Substrate Plant 5-Lipoxygenases Behave like 11-Lipoxygenases

Zadelhoff

Veldink

Vliegenthart

1998

Biochemical and Biophysical Research Communications

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“…Its chemical structure was confirmed by HPLC analysis, including chiral-phase HPLC [12], ultraviolet and infrared spectroscopy and GC/MS.…”

Section: Preparationsmentioning

confidence: 99%

“…Chiral-phase HPLC was carried out on a Bakerbond chiral-phase HPLC column (dinitrobenzoylphenylglycine ionicly linked to silica gel as stationary phase) [12]. Protein concentration of the lipoxygenase preparation was determined according to [19].…”

Section: Miscellaneousmentioning

confidence: 99%

Formation of lipoxin B by the pure reticulocyte lipoxygenase via sequential oxygenation of the substrate

Kühn

Wiesner

Alder

et al. 1987

European Journal of Biochemistry

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The pure reticulocyte lipoxygenase converts 1 5LS-hydroxy-5,8,1 1,13(2,Z,Z,E)-icosatetraenoic acid (1 5~~-HETE) methyl ester to a complex mixture of products containing 5~, , 1 4~~, 1 5~~-trihydro(pero)xy-6E,8Z,lOE,12E-icosatetraenoate methyl ester (lipoxin B methyl ester), 5~, , l SL,-D~H(P)ETE methyl ester and four 8 , 1 5~~-DiH(P)ETE methyl ester isomers [DiH(P)ETE = dihydro(pero)xy-icosatetraenoic acid].After a short incubation period (15 min) 5os,15~,-DiH(P)ETE methyl ester was found to be the main product, whereas after a 3-h incubation lipoxin B methyl ester was the predominant product. The reaction shows a remarkable stereoselectivity since only small amounts of other trihydroxy tetraenes are formed. Anaerobiosis, heat inactivation of the enzyme, or incubation in the presence of lipoxygenase inhibitors (icosatetraynoic acid, nordihydroguaiaretic acid) completely abolished the reaction. The complete steric structure of the major tetraene product (lipoxin B methyl ester) was established by ultraviolet spectroscopy, HPLC on four different types of columns, gas chromatography/mass spectrometry, gas/liquid chromatography of the ozonolysis fragments of the menthoxycarbonyl derivatives, and by 400-MHz 'H-NMR.Atmospheric oxygen was incorporated at carbon-5 and carbon-14 into the major product. 5~~, 1 5~~-DiH(P)ETE methyl ester was shown to be an intermediate in the synthesis. Lipoxin B was also formed during the oxygenation of arachidonic acid, 1 SL,-HETE and 5~~, l SL,-D~HETE. The results presented here indicate that lipoxin B can be formed by pure lipoxygenases via a sequential oxygenation of arachidonic acid or its hydro(pero)xy derivatives.

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“…Individual isomers, separated by SP-HPLC according to position of oxidation and diene configuration, were subsequently analyzed for R and S isomeric composition by CP-HPLC as described by Kuhn et al. (16).…”

Section: Hplc Separationsmentioning

confidence: 99%

Hydroperoxide Lyase and Other Hydroperoxide-Metabolizing Activity in Tissues of Soybean, Glycine max

Gardner¹,

Weisleder²,

Plattner³

1991

Plant Physiol.

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Hydroperoxide lyase (HPLS) activity in soybean (Glycine max) seed/seedlings, leaves, and chloroplasts of leaves required detergent solubilization for maximum in vitro activity. On a per milligram of protein basis, more HPLS activity was found in leaves, especially chloroplasts, than in seeds or seedlings. The total yield of hexanal from 13(S)-hydroperoxy-cis-9,trans-1 1-octadecadienoic acid (13S-HPOD) from leaf or chloroplast preparations was 58 and 66 to 85%, respectively. Because of significant competing hydroperoxide-metabolizing activities from other enzymes in seed/seedling preparations, the hexanal yields from this source were lower (36-56%). Some of the products identified from the seed or seedling preparations indicated that the competing activity was mainly due to both a hydroperoxide peroxygenase and reactions catalyzed by lipoxygenase. Different HPLS isozyme compositions in the seed/seedling versus the leaf/chloroplast preparations were indicated by differences in the activity as a function of pH, the Km values, relative V .. with 13S-HPOD and 13(S)-hydroperoxy-cis-9,trans-1 ,cis-15-octadecatrienoic acid (13S-HPOT), and the specificity with different substrates. With regard to the latter, both seed/seedling and chloroplast HPLS utilized the 13S-HPOD and 13S-HPOT substrates, but only seeds/ seedlings were capable of metabolizing 9(S)-hydroperoxy-trans-10,cis-12-octadecadienoic acid into 9-oxononanoic acid, isomenc nonenals, and 4-hydroxynonenal. From 13S-HPOD and 13S-HPOT, the products were identified as 12-oxo-cis-9-dodecenoic acid, as well as hexanal from 13S-HPOD and cis-3-hexenal from 13S-HPOT. In seed preparations, there was partial isomerization of the cis-3 or cis-9 into trans-2 or trans-10 double bonds, respectively.HPLS', an enzyme that cleaves hydroperoxides of polyunsaturated fatty acids into both aldehyde and w-oxoacid fragments, occurs widely in plants. Depending on substrate specificity of HPLS isoenzymes for isomeric hydroperoxides of linoleic or linolenic acids, the enzyme produces either green/ ' Abbreviations: HPLS, hydroperoxide lyase; CP-HPLC, chiralphase HPLC; El, electron impact; OTMS, trimethylsilyloxy; rac-HPOD, racemic cis,trans-and trans,trans-diene linoleic acid hydroperoxides; rac-HPOT, racemic cis,cis,trans-triene linolenic acid hydroperoxides; 9S-HPOD, 9(S)-hydroperoxy-trans-10,cis-12-octadecadienoic acid; 13S-HPOD, 13(S)-hydroperoxy-cis-9,trans-11-octadecadienoic acid; 12S-HPOT, 12(S)-hydroperoxy-cis-9,trans-13,cis-1 5-octadecatrienoic acid; 1 3S-HPOT, 1 3(S)-hydroperoxy-cis-9,trans-11 ,cis-5-octadecatrienoic acid; SP-HPLC, straight-phase HPLC; Triton X-1 OOR, Triton X-100 reduced; m/z, mass to charge ratio. beany/grassy odors (hexanal or cis-3-hexenal) from 1 3-hydroperoxides or cucumber/pear odors (cis-3-nonenal or cis-3,cis-6 nonadienal) from 9-hydroperoxides (reviewed in refs. 4 and 15). The corresponding w-oxoacid fragments, 12-oxo-cis-9-dodecenoic acid and 9-oxononanoic acid, have no known function, except the former isomerizes into 12-oxo-trans-10-dodecenoi...

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Analysis of the stereochemistry of lipoxygenase-derived hydroxypolyenoic fatty acids by means of chiral phase high-pressure liquid chromatography

Cited by 123 publications

References 16 publications

With Anandamide as Substrate Plant 5-Lipoxygenases Behave like 11-Lipoxygenases

With Anandamide as Substrate Plant 5-Lipoxygenases Behave like 11-Lipoxygenases

Formation of lipoxin B by the pure reticulocyte lipoxygenase via sequential oxygenation of the substrate

Hydroperoxide Lyase and Other Hydroperoxide-Metabolizing Activity in Tissues of Soybean, Glycine max

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