Abstract
We have prepared Schiff base adducts of the core aldehydes of phosphatidylcholine and aminophospholipids, free amino acids, and myoglobin. The Schiff bases of the ethanolamine and serine glycerophospholipids were obtained by reacting sn-1-palmitoyl(stearoyl)-2-[9-oxo]nonanoyl-glycerophosphocholine (PC-Ald) with a twofold excess of the aminophospholipid in chloroform/methanol 2∶1 (vol/vol) for 18 h at room temperature. The Schiff bases of the amino acids and myoglobin were obtained by reacting the aldehyde with an excess of isoleucine, valine, lysine, methyl ester lysine and myoglobin in aqueous methanol for 18 h at room temperature. Prior to isolation, the Schiff bases were reduced with sodium cyanoborohydride in methanol for 30 min at 4°C. The reaction products were characterized by normal-phase high-performance liquid chromatography and on-line mass spectrometry with electrospray ionization. The amino acids and aminophospholipids yielded single adducts. A double adduct was obtained for myoglobin, which theoretically could have accepted up to 23 PC-Ald groups. The yields of the products ranged from 12 to 44% for the aminophospholipids and from 15–57% for the amino acids, while the Schiff base of the myoglobin was estimated at 5% level. The new compounds are used as reference standards for the detection of high molecular weight Schiff bases in lipid extracts of natural products.
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Abbreviations
- GPC:
-
glycerophosphocholine
- PE:
-
glycerophosphoethanolamine
- GPS:
-
glycerophosphoserine
- HPLC:
-
high-performance liquid chromatography
- LC/ESI/MS:
-
liquid chromatography/electrospray ionization/mass spectrometry
- PAF:
-
platelet-activating factor
- PC:
-
phosphatidylcholine
- PC-ALD:
-
1-palmitoyl(stearoyl)-2-[9-oxolnonanoyl GPC
- PE:
-
phosphatidylethanolamine
- PS:
-
phosphatidylserine
- TLC:
-
thin-layer chromatography
References
Esterbauer, H., Zollner, H., and Schaur, R.J. (1989) Aldehydes Formed by Lipid Peroxidation: Mechanism of Formation, Occurrence and Determination, in Membrane Lipid Oxidation (Pelfrey, C., ed.) pp. 239–268, CRC Press, Boea Raton.
Crawford, D.L., Yu, T.C., and Sinhuber, R.O. (1966) Reaction of Malonaldehyde with Glycine, J. Agr. Food Chem. 14, 182–184.
Sawicki, E., Stanley, T.W., and Johnson, H. (1963) Comparison of Spectrophotometric and Spectrophotofluorometric Methods for the Determination of Malonaldehyde, Anal. Chem. 35, 199–205.
Chio, K.S., and Tappel, A.L. (1969) Synthesis and Characterization of the Fluorescent Products Derived from Malonaldehyde and Amino Acids. Biochemistry 8, 2821–2827.
Nair, V., Vietti, D.E., and Cooper, C.S. (1981) Degenerative Chemistry of Malonadehyde. Structure, Stereochemistry, and Kinetics of Formation of Enaminals from Reaction with Amino Acids, J. Am. Chem. Soc., 103, 3030–3036.
Chaudhary, A.K., Nokubo, M., Reddy, G.R., Yeola, S.N., Morrow, J.D., Blair, I.A., and Marnett, L.J. (1994) Detection of Endogenous Malonaldehyde-Deoxyguanosine Adducts in Human Liver. Science 265, 1580–1582.
Nadkarni, D.V., and Sayre, L.M. (1995) Structural Definition of Early Lysine and Histidine Adduction Chemistry of 4-Hydroxynonenal, Chem. Res. Toxicol. 8, 284–291.
Bruenner, B.A., Jones, A.D., and German, J.B. (1995) Direct Characterization of Protein Adducts of the Lipid Peroxidation Product 4-Hydroxy-2-Nonenal Using Electrospray Mass Spectrometry. Chem. Res. Toxicol. 8, 552–559.
Szweda, L.I., Uchida, K., Tsai, L., and Stadtman, E.R. (1993) Inactivation of Glucose-6-phosphate Dehydrogenase b, 4-Hydroxy-2-nonenal. Selective Modification of an Active-Site Lysine, J. Biol. Chem. 268, 3342–3347.
Uchida, K., Toyokuni, S., Nishikawa, K., Kawakishi, S., Oda, H., Hiai, H., and Stadtman, E.R. (1994) Michael Addition-type 4-Hydroxy-2-nonenal Adducts in Modified Low-Density Lipoproteins: Markers for Atherosclerosis, Biochemistry 33, 12487–12494.
Bruenner, B.A., Jones, A.D., and German, J.B. (1994) Maximum Entropy Deconvolution of Heterogeneity in Protein Modification: Protein Adducts of 4-Hydroxy-2-nenenal, Rapid Commun. Mass Spectrom. 8, 509–512.
Wang, C.-J., and Tai, H.-H. (1990) A Facile Synthesis of an Aldehydic Analog of Platelet-Activating Factor and Its Use in the Production of Specific Antibodies, Chem. Phys. Lipids 55, 265–273.
Kamido, H., Kuksis, A., Marai, L., and Myher, J.J. (1992) Identification of Cholesterol-Bound Aldehydes in Copper-Oxidized Low Density Lipoprotein. FEBS Lett. 304, 269–272.
Kamido, H., Kuksis, A., Marai, L., and Myher, J.J. (1995) Lipid Ester-Bound Aldehydes Among Copper-Catalyzed Peroxidation Products of Human Plasma Lipoproteins. J. Lipid Res. 36, 1876–1886.
Biemann, K. (1992) Mass Spectrometry of Peptides and Proteins, Ann. Rev. Biochem. 61, 977–1010.
Privett, O.S., and Blank, M.L. (1961) A New Method for the Structural Analysis of Component Mono-, Di- and Triglycerides, J. Lipid Res 2, 37–44.
Ravandi, A., Kuksis, A., Myher, J.J., and Marai, L. (1995) Determination of Lipid Ester Ozonides and Core Aldehydes by High-Performance Liquid Chromatography with On-Line Mass Spectrometry, J. Biochem. Biophys. Methods 30, 271–285.
Myher, J.J., Kuksis, A., Ravandi, A., and Cocks, N. (1994) Normal-Phase Liquid Chromatography/Mass Spectrometry with Electrospray for Sensitive Detection of Oxygenated Glycerophospholipids, INFORM 5, 478–479. Abstr. No E.
Becart, J., Chevalier, C., and Biesse, J.P. (1990) Quantitative Analysis of Phospholipids by HPLC with a Light-Scattering Evaporating Detector—Application to Raw Materials of Cosmetic Use, J. High Resol. Chromatogr. 13, 126–129.
Kim, H.-Y., Wang, T.-C.L., and Ma, Y.-C. (1994) Liquid Chromatography/Mass Spectrometry of Phospholipids Using Electrospray Ionization. Anal. Chem. 66, 3977–3982.
Stapelfeldt, H., and Skibsted, L.H. (1996) Kinetics of Formation of Fluorescent Products from Hexanal and l-Lysine in a Two-Phase System, Lipids 31, 1125–1132.
Jain, S.K., and Shohet, S.B. (1984) A Novel Phospholipid in Irreversibly Sickled Cells: Evidence for in vivo Peroxidative Membrane Damage in Sickle Cell Disease, Blood 63, 362–367.
Ravandi, A., Kuksis, A., Marai, L., and Myher, J.J. (1995) Preparation and Characterization of Glucosylated Aminoglycerophospholipids, Lipids 30, 885–891.
Pamplona, R., Bellmunt, M.J., Portero, M., Riba, D., and Prat, J. (1995) Chromatographic Evidence for Amadori Product Formation in Rat Liver Aminophospholipids, Life Sciences 57, 873–879.
McMillen, D.A., Volwerk, J.J., Ohishi, J.-I., Erion, M., Keana, J.F.W., Jost, P.C., and Griffith, O.H. (1986) Identifying Regions of Membrane Proteins in Contact with Phospholipid Head Groups: Covalent Attachment of a New Class of Aldehyde Lipid Labels to Cytochrome C Oxidase, Biochemistry 25, 182–193.
Le Blanc, J.C.Y., Siu, K.W.M., and Guevremont, R. (1994) Electrospray Mass Spectrometric Study of Protein-Ketone Equilibria in Solution, Anal. Chem. 66, 3289–3296.
Steinbrecher, U.P. (1987) Oxidation of Human Low Density Lipoprotein Results in Derivatization of Lysine Residues of Apolipoprotein B by Lipid Peroxide Decomposition Products, J. Biol. Chem. 262, 3603–3608.
Haberland, M.E., Fong, D., and Cheng, L. (1988) Malonaldehyde-Altered Protein Occurs in Atheroma of Watanabe Heritable Hyperlipidemic Rabbits, Science, 241, 215–218.
Juergens, G., Chen, O., Esterbauer, H., Mair, S., Ledinski, G., and Dinges, H.P. (1993) Immunostaining of Human Autopsy Aortas with Antibodies to Modified Apolipoprotein B and Apoprotein (A), Arterioscler. Thromb. 13, 1689–1699.
Malle, E., Ibovnik, A., Leis, H.J., Kostner, G.M., Verhallen, P.F.J., and Sattler, W. (1995) Lysine Modification of LDL or Lipoprotein (a) by 4-Hydroxynonenal or Malonaldehyde Decreases Platelet Aggregability and Eicosanoid Formation. Arteriscler. Thromb. Vasc. Biol. 15, 377–384.
Jain, S.K., Yip, R., Hoesch, R.M., Pramanik, A.K., Dallman, P.R., and Shohet, S.B. (1983) Evidence for Peroxidative Damage to the Erythrocyte Membrane in Iron Deficiency. Am. J. Clin. Nutr. 37, 26–30.
Jain, S.K., and Hochstein, P. (1980) Polymerization of Membrane Components in Ageing Red Blood Cells. Biochem. Biophys. Res. Commun. 92, 247–254.
Horkko, S., Miller, E., Dudl, E., Reaven, P., Curtiss, L.K., Zvaifler, N.J., Terkeltaub, R., Pierangeli, S.S., Ware Branch, D., Palinski, W., and Witzum, J.J. (1996). Antiphospholipid Antibodies are Directed Against Epitopes of Oxidized Phospholipids. Recognition of Cardiolipin by Monoclonal Antibodies to Epitopes of Oxidized Low Density Lipoprotein, J. Clin. Invest. 98, 815–825.
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Based on presentation at the AOCS Annual Meeting & Expo in Indianapolis, Indiana, April 28–May 1, 1996.
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Ravandi, A., Kuksis, A., Shaikh, N. et al. Preparation of schiff base adducts of phosphatidylcholine core aldehydes and aminophospholipids, amino acids, and myoglobin. Lipids 32, 989–1001 (1997). https://doi.org/10.1007/s11745-997-0129-6
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DOI: https://doi.org/10.1007/s11745-997-0129-6