Advertisement

Analytical and Bioanalytical Chemistry

, Volume 407, Issue 17, pp 5161–5173 | Cite as

Identification of carbonylated lipids from different phospholipid classes by shotgun and LC-MS lipidomics

  • Zhixu Ni
  • Ivana Milic
  • Maria FedorovaEmail author
Research Paper
Part of the following topical collections:
  1. Lipidomics

Abstract

Oxidized lipids play a significant role in the pathogenesis of numerous oxidative stress-related human disorders, such as atherosclerosis, obesity, inflammation, and autoimmune diseases. Lipid peroxidation, induced by reactive oxygen and nitrogen species, yields a high variety of modified lipids. Among them, carbonylated lipid peroxidation products (oxoLPP), formed by oxidation of the fatty acid moiety yielding aldehydes or ketones (carbonyl groups), are electrophilic compounds that are able to modify nucleophilic substrates like proteins, nucleic acid, and aminophospholipids. Some carbonylated phosphatidylcholines possess even pro-inflammatory activities. However, little is known about oxoLPP derived from other phospholipid (PL) classes. Here, we present a new analytical strategy based on the mass spectrometry (MS) of PL-oxoLPP derivatized with 7-(diethylamino)coumarin-3-carbohydrazide (CHH). Shotgun MS revealed many oxoLPP derived from in vitro oxidized glycerophosphatidylglycerols (PG, 31), glycerophosphatidylcholine (PC, 23), glycerophosphatidylethanolamine (PE, 34), glycerophosphatidylserines (PS, 7), glycerophosphatidic acids (PA, 17), and phosphatidylinositiolphosphates (PIP, 6) vesicles. This data were used to optimize LipidXplorer-assisted identification, and a python-based post-processing script was developed to increase both throughput and accuracy. When applied to full lipid extracts from rat primary cardiomyocytes treated with peroxynitrite donor SIN-1, ten PL-bound oxoLPP were unambiguously identified by LC-MS, including two PC-, two PE-, one PG-, two PS-, and three PA-derived species. Some of the well-known carbonylated PC were detected, while most PL-oxoLPP were shown for the first time.

Graphical Abstract

Overview of analytical and bioinformatics approach for detection and identification of carbonylated phospholipids.

Keywords

7-(Diethylamino)coumarin-3-carbohydrazide Carbonylated lipids LC-MS Lipid peroxidation products LipidXplorer Shotgun lipidomics 

Abbreviations

AA

Arachidonic acid

CHH

7-(Diethylamino)coumarin-3-carbohydrazide

CID

Collision-induced dissociation

DAG

Diacylglycerols

DBE

Double bond equivalents

DDA

Data-dependent acquisition

DHA

Docosahexaenoic acid

ESI

Electrospray

FA

Fatty acid

GPF

Gas phase fractionation

HG

Head group

HOOA-PC

1-Palmitoyl-2-(5-hydroxy-8-oxo-oct-6-enoyl)-glycerophosphatidylcholine

LA

Linoleic acid

LC-MS

Liquid chromatography coupled to mass spectrometry

LPP

Lipid peroxidation products

MFQL

Molecular fragmentation query language

MS

Mass spectrometry

MS/MS

Tandem mass spectrometry

MTBE

Methyl-tert-butyl ether

nCE

Normalized collision energy

OA

Oleic acid

OAP

Oxygen addition products

OCP

Oxygen cleavage products

oxoLPP

Carbonylated lipid peroxidation products

oxPC

Oxidized phosphatidylcholine

PA

Glycerophosphatidic acids

PAPC

1-Palmitoyl-2-arachidonyl-glycerophosphatidylcholine

PAPE

1-Palmitoyl-2-arachidonoyl-glycerophosphatidylethanolamine

PAPG

1-Palmitoyl-2-arachidonoyl-glycerophosphatidylglycerol

PC

Glycerophosphatidylcholine

PDPC

1-Palmitoyl-2-docosahexaenoyl-glycerophosphatidylcholine

PDPE

1-Palmitoyl-2-docosahexaenoyl-glycerophosphatidylethanolamine

PE

Glycerophosphatidylethanolamine

PI

Glycerophosphatidylinositols

PIP

Phosphatidylinositiolphosphates

PL

Glycerophospholipids

PLPA

1-Palmitoyl-2-linoleoyl-glycerophosphatidic acid

PLPC

1-Palmitoyl-2-linoleoyl-glycerophosphatidylcholine

PLPE

1-Palmitoyl-2-linoleoyl-glycerophosphatidylethanolamine

PLPG

1-Palmitoyl-2-linoleoyl-glycerophosphatidylglycerol

PLPS

1-Palmitoyl-2-linoleoyl-glycerophosphatidylserine

POPA

1-Palmitoyl-2-oleoyl-glycerophosphatidic acid

POPC

1-Palmitoyl-2-oleoyl-glycerophosphatidylcholine

POPE

1-Palmitoyl-2-oleoyl-glycerophosphatidylethanolamine

POPG

1-Palmitoyl-2-oleoyl-glycerophosphatidylglycerol

POPS

1-Palmitoyl-2-oleoyl-glycerophosphatidylserine

POVPC

1-Palmitoyl-2-(5-oxo-valeryl)-glycerophosphatidylcholine

PS

Glycerophosphatidylserines

PUFA

Polyunsaturated fatty acid

RNS

Reactive nitrogen species

ROS

Reactive oxygen species

RPC

Reversed phase chromatography

SAPC

1-Stearoyl-2-arachidonoylglycerophosphatidylcholine

SAPIP

1-Stearoyl-2-arachidonoyl-gylcerophosphatidylinositolphosphate

SIN-1

3-Morpholinosydnonimine

SLPIP

1-Stearoyl-2-linoleoyl-glycerophosphatidylinositolphosphate

Notes

Acknowledgments

The authors are thankful to Prof. Ralf Hoffmann (Institute of Bioanalytical Chemistry, University of Leipzig) for providing access to his laboratory and mass spectrometers. Financial support from European Regional Development Fund (ERDF, European Union and Free State Saxony; 100146238 and 100121468 to M.F) and a stipend to I.M. provided by Universität Leipzig are gratefully acknowledged.

Supplementary material

216_2015_8536_MOESM1_ESM.pdf (475 kb)
ESM 1 (PDF 474 kb)

References

  1. 1.
    Dowhan W, Bogdanov M, Mileykovskaya E (2008) Chapter 1—functional roles of lipids in membranes. In: Deve V (ed) Biochemistry of lipids, lipoproteins and membranes, 5th edn. Elsevier, San DiegoGoogle Scholar
  2. 2.
    Yeagle PL (1989) Lipid regulation of cell membrane structure and function. FASEB J 3(7):1833–1842Google Scholar
  3. 3.
    Wymann MP, Schneiter R (2008) Lipid signalling in disease. Nat Rev Mol Cell Biol 9(2):162–176CrossRefGoogle Scholar
  4. 4.
    Doria ML, Cotrim CZ, Simoes C, Macedo B, Domingues P, Domingues MR, Helguero LA (2013) Lipidomic analysis of phospholipids from human mammary epithelial and breast cancer cell lines. J Cell Physiol 228(2):457–468CrossRefGoogle Scholar
  5. 5.
    Han X, Gross RW (2003) Global analyses of cellular lipidomes directly from crude extracts of biological samples by ESI mass spectrometry: a bridge to lipidomics. J Lipid Res 44(6):1071–1079CrossRefGoogle Scholar
  6. 6.
    Schwudke D, Schuhmann K, Herzog R, Bornstein SR, Shevchenko A (2011) Shotgun lipidomics on high resolution mass spectrometers. Cold Spring Harb Perspect Biol 3(9):a004614CrossRefGoogle Scholar
  7. 7.
    Davi G, Falco A, Patrono C (2005) Lipid peroxidation in diabetes mellitus. Antioxid Redox Signal 7(1–2):256–268CrossRefGoogle Scholar
  8. 8.
    Reed TT (2011) Lipid peroxidation and neurodegenerative disease. Free Radic Biol Med 51(7):1302–1319CrossRefGoogle Scholar
  9. 9.
    Yagi K (1987) Lipid peroxides and human diseases. Chem Phys Lipids 45(2–4):337–351CrossRefGoogle Scholar
  10. 10.
    Sultana R, Perluigi M, Allan Butterfield D (2013) Lipid peroxidation triggers neurodegeneration: a redox proteomics view into the Alzheimer disease brain. Free Radic Biol Med 62:157–169CrossRefGoogle Scholar
  11. 11.
    Shao B, Heinecke JW (2009) HDL, lipid peroxidation, and atherosclerosis. J Lipid Res 50(4):599–601CrossRefGoogle Scholar
  12. 12.
    Esterbauer H, Wag G, Puhl H (1993) Lipid peroxidation and its role in atherosclerosis. Br Med Bull 49(3):566–576Google Scholar
  13. 13.
    Watson AD (2006) Thematic review series: systems biology approaches to metabolic and cardiovascular disorders. Lipidomics: a global approach to lipid analysis in biological systems. J Lipid Res 47(10):2101–2111CrossRefGoogle Scholar
  14. 14.
    Olusi SO (2002) Obesity is an independent risk factor for plasma lipid peroxidation and depletion of erythrocyte cytoprotectic enzymes in humans. Int J Obes Relat Metab Disord 26(9):1159–1164CrossRefGoogle Scholar
  15. 15.
    Basu S, Riserus U, Turpeinen A, Vessby B (2000) Conjugated linoleic acid induces lipid peroxidation in men with abdominal obesity. Clin Sci (Lond) 99(6):511–516CrossRefGoogle Scholar
  16. 16.
    Zhang R, Brennan M-L, Shen Z, MacPherson JC, Schmitt D, Molenda CE, Hazen SL (2002) Myeloperoxidase functions as a major enzymatic catalyst for initiation of lipid peroxidation at sites of inflammation. J Biol Chem 277(48):46116–46122CrossRefGoogle Scholar
  17. 17.
    Minoguchi K, Yokoe T, Tanaka A, Ohta S, Hirano T, Yoshino G, O’Donnell CP, Adachi M (2006) Association between lipid peroxidation and inflammation in obstructive sleep apnoea. Eur Respir J 28(2):378–385CrossRefGoogle Scholar
  18. 18.
    Michel P, Eggert W, Albrecht-Nebe H, Grune T (1997) Increased lipid peroxidation in children with autoimmune diseases. Acta Paediatr 86(6):609–612CrossRefGoogle Scholar
  19. 19.
    Wang G, Konig R, Ansari GA, Khan MF (2008) Lipid peroxidation-derived aldehyde-protein adducts contribute to trichloroethene-mediated autoimmunity via activation of CD4+ T cells. Free Radic Biol Med 44(7):1475–1482CrossRefGoogle Scholar
  20. 20.
    Niki E, Yoshida Y, Saito Y, Noguchi N (2005) Lipid peroxidation: mechanisms, inhibition, and biological effects. Biochem Biophys Res Commun 338(1):668–676CrossRefGoogle Scholar
  21. 21.
    Reis A, Spickett CM (2012) Chemistry of phospholipid oxidation. BBA-Biomembr 1818(10):2374–2387CrossRefGoogle Scholar
  22. 22.
    Milic I, Fedorova M, Teuber K, Schiller J, Hoffmann R (2012) Characterization of oxidation products from 1-palmitoyl-2-linoleoyl-sn-glycerophosphatidylcholine in aqueous solutions and their reactions with cysteine, histidine and lysine residues. Chem Phys Lipids 165(2):186–196CrossRefGoogle Scholar
  23. 23.
    Rauniyar N, Prokai L (2009) Detection and identification of 4-hydroxy-2-nonenal Schiff-base adducts along with products of Michael addition using data-dependent neutral loss-driven MS(3) acquisition: method evaluation through an in vitro study on cytochrome c oxidase modifications. Proteomics 9(22):5188–5193CrossRefGoogle Scholar
  24. 24.
    Rikans LE, Hornbrook KR (1997) Lipid peroxidation, antioxidant protection and aging. BBA-Mol Basis Dis 1362(2–3):116–127CrossRefGoogle Scholar
  25. 25.
    Spiteller G (2001) Lipid peroxidation in aging and age-dependent diseases. Exp Gerontol 36(9):1425–1457CrossRefGoogle Scholar
  26. 26.
    Montine TJ, Neely MD, Quinn JF, Beal MF, Markesbery WR, Roberts LJ, Morrow JD (2002) Lipid peroxidation in aging brain and Alzheimer’s disease. Free Radic Biol Med 33(5):620–626CrossRefGoogle Scholar
  27. 27.
    Guo L, Chen Z, Amarnath V, Davies SS (2012) Identification of novel bioactive aldehyde-modified phosphatidylethanolamines formed by lipid peroxidation. Free Radic Biol Med 53(6):1226–1238CrossRefGoogle Scholar
  28. 28.
    Annibal A, Schubert K, Wagner U, Hoffmann R, Schiller J, Fedorova M (2014) New covalent modifications of phosphatidylethanolamine by alkanals: mass spectrometry based structural characterization and biological effects. J Mass Spectrom 49(7):557–569CrossRefGoogle Scholar
  29. 29.
    Bacot S, Bernoud-Hubac N, Baddas N, Chantegrel B, Deshayes C, Doutheau A, Lagarde M, Guichardant M (2003) Covalent binding of hydroxy-alkenals 4-HDDE, 4-HHE, and 4-HNE to ethanolamine phospholipid subclasses. J Lipid Res 44(5):917–926CrossRefGoogle Scholar
  30. 30.
    Berliner JA, Leitinger N, Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis. J Lipid Res 50(Suppl):S207–S212Google Scholar
  31. 31.
    Berliner JA, Watson AD (2005) A role for oxidized phospholipids in atherosclerosis. N Engl J Med 353(1):9–11CrossRefGoogle Scholar
  32. 32.
    Stemmer U, Hermetter A (2012) Protein modification by aldehydophospholipids and its functional consequences. BBA-Biomembr 1818(10):2436–2445CrossRefGoogle Scholar
  33. 33.
    Loidl A, Sevcsik E, Riesenhuber G, Deigner H-P, Hermetter A (2003) Oxidized phospholipids in minimally modified low density lipoprotein induce apoptotic signaling via activation of acid sphingomyelinase in arterial smooth muscle cells. J Biol Chem 278(35):32921–32928CrossRefGoogle Scholar
  34. 34.
    Fruhwirth GO, Moumtzi A, Loidl A, Ingolic E, Hermetter A (2006) The oxidized phospholipids POVPC and PGPC inhibit growth and induce apoptosis in vascular smooth muscle cells. Biochim Biophys Acta 1761(9):1060–1069CrossRefGoogle Scholar
  35. 35.
    Podrez EA, Poliakov E, Shen Z, Zhang R, Deng Y, Sun M, Finton PJ, Shan L, Febbraio M, Hajjar DP, Silverstein RL, Hoff HF, Salomon RG, Hazen SL (2002) A novel family of atherogenic oxidized phospholipids promotes macrophage foam cell formation via the scavenger receptor CD36 and is enriched in atherosclerotic lesions. J Biol Chem 277(41):38517–38523CrossRefGoogle Scholar
  36. 36.
    Milic I, Fedorova M (2015) Derivatization and detection of small aliphatic and lipid-bound carbonylated lipid peroxidation products by ESI-MS. Methods Mol Biol 1208:3–20CrossRefGoogle Scholar
  37. 37.
    Milic I, Hoffmann R, Fedorova M (2013) Simultaneous detection of low and high molecular weight carbonylated compounds derived from lipid peroxidation by electrospray ionization-tandem mass spectrometry. Anal Chem 85(1):156–162CrossRefGoogle Scholar
  38. 38.
    Herzog R, Schuhmann K, Schwudke D, Sampaio JL, Bornstein SR, Schroeder M, Shevchenko A (2012) LipidXplorer: a software for consensual cross-platform lipidomics. PLoS ONE 7(1):e29851CrossRefGoogle Scholar
  39. 39.
    Herzog R, Schwudke D, Schuhmann K, Sampaio JL, Bornstein SR, Schroeder M, Shevchenko A (2011) A novel informatics concept for high-throughput shotgun lipidomics based on the molecular fragmentation query language. Genome Biol 12(1):R8CrossRefGoogle Scholar
  40. 40.
    Matyash V, Liebisch G, Kurzchalia TV, Shevchenko A, Schwudke D (2008) Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics. J Lipid Res 49(5):1137–1146CrossRefGoogle Scholar
  41. 41.
    Chambers MC, Maclean B, Burke R, Amodei D, Ruderman DL, Neumann S, Gatto L, Fischer B, Pratt B, Egertson J, Hoff K, Kessner D, Tasman N, Shulman N, Frewen B, Baker TA, Brusniak M-Y, Paulse C, Creasy D, Flashner L, Kani K, Moulding C, Seymour SL, Nuwaysir LM, Lefebvre B, Kuhlmann F, Roark J, Rainer P, Detlev S, Hemenway T, Huhmer A, Langridge J, Connolly B, Chadick T, Holly K, Eckels J, Deutsch EW, Moritz RL, Katz JE, Agus DB, MacCoss M, Tabb DL, Mallick P (2012) A cross-platform toolkit for mass spectrometry and proteomics. Nat Biotechnol 30(10):918–920CrossRefGoogle Scholar
  42. 42.
    Reis A, Domingues P, Ferrer-Correia AJ, Domingues MR (2004) Fragmentation study of short-chain products derived from oxidation of diacylphosphatidylcholines by electrospray tandem mass spectrometry: identification of novel short-chain products. Rapid Commun Mass Spectrom 18(23):2849–2858CrossRefGoogle Scholar
  43. 43.
    Spickett CM, Reis A, Pitt AR (2011) Identification of oxidized phospholipids by electrospray ionization mass spectrometry and LC-MS using a QQLIT instrument. Free Radic Biol Med 51(12):2133–2149CrossRefGoogle Scholar
  44. 44.
    Reis A, Domingues MRM, Amado FML, Ferrer-Correia AJ, Domingues P (2007) Radical peroxidation of palmitoyl-lineloyl-glycerophosphocholine liposomes: identification of long-chain oxidised products by liquid chromatography–tandem mass spectrometry. J Chromatogr B 855(2):186–199CrossRefGoogle Scholar
  45. 45.
    Mak TD, Laiakis EC, Goudarzi M, Fornace AJ Jr (2014) MetaboLyzer: a novel statistical workflow for analyzing postprocessed LC-MS metabolomics data. Anal Chem 86(1):506–513CrossRefGoogle Scholar
  46. 46.
    Kind T, Okazaki Y, Saito K, Fiehn O (2014) LipidBlast templates as flexible tools for creating new in-silico tandem mass spectral libraries. Anal Chem 86(22):11024–11027CrossRefGoogle Scholar
  47. 47.
    Husen P, Tarasov K, Katafiasz M, Sokol E, Vogt J, Baumgart J, Nitsch R, Ekroos K, Ejsing CS (2013) Analysis of lipid experiments (ALEX): a software framework for analysis of high-resolution shotgun lipidomics data. PLoS ONE 8(11):e79736CrossRefGoogle Scholar
  48. 48.
    Song H, Hsu F-F, Ladenson J, Turk J (2007) Algorithm for processing raw mass spectrometric data to identify and quantitate complex lipid molecular species in mixtures by data-dependent scanning and fragment ion database searching. J Am Soc Mass Spectrom 18(10):1848–1858CrossRefGoogle Scholar
  49. 49.
    Hubner G, Crone C, Lindner B (2009) lipID—a software tool for automated assignment of lipids in mass spectra. J Mass Spectrom 44(12):1676–1683Google Scholar
  50. 50.
    Strohalm M, Kavan D, Novak P, Volny M, Havlicek V (2010) mMass 3: a cross-platform software environment for precise analysis of mass spectrometric data. Anal Chem 82(11):4648–4651CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Institute of Bioanalytical Chemistry, Faculty of Chemistry and MineralogyUniversität LeipzigLeipzigGermany
  2. 2.Center for Biotechnology and BiomedicineUniversität LeipzigLeipzigGermany

Personalised recommendations