Analytical and Bioanalytical Chemistry

, Volume 407, Issue 20, pp 5965–5976 | Cite as

Differential proteomic analysis of mouse macrophages exposed to adsorbate-loaded heavy fuel oil derived combustion particles using an automated sample-preparation workflow

  • Tamara Kanashova
  • Oliver Popp
  • Jürgen Orasche
  • Erwin Karg
  • Horst Harndorf
  • Benjamin Stengel
  • Martin Sklorz
  • Thorsten Streibel
  • Ralf Zimmermann
  • Gunnar Dittmar
Research Paper
Part of the following topical collections:
  1. Aerosols and Health

Abstract

Ship diesel combustion particles are known to cause broad cytotoxic effects and thereby strongly impact human health. Particles from heavy fuel oil (HFO) operated ships are considered as particularly dangerous. However, little is known about the relevant components of the ship emission particles. In particular, it is interesting to know if the particle cores, consisting of soot and metal oxides, or the adsorbate layers, consisting of semi- and low-volatile organic compounds and salts, are more relevant. We therefore sought to relate the adsorbates and the core composition of HFO combustion particles to the early cellular responses, allowing for the development of measures that counteract their detrimental effects. Hence, the semi-volatile coating of HFO-operated ship diesel engine particles was removed by stepwise thermal stripping using different temperatures. RAW 264.7 macrophages were exposed to native and thermally stripped particles in submersed culture. Proteomic changes were monitored by two different quantitative mass spectrometry approaches, stable isotope labeling by amino acids in cell culture (SILAC) and dimethyl labeling. Our data revealed that cells reacted differently to native or stripped HFO combustion particles. Cells exposed to thermally stripped particles showed a very differential reaction with respect to the composition of the individual chemical load of the particle. The cellular reactions of the HFO particles included reaction to oxidative stress, reorganization of the cytoskeleton and changes in endocytosis. Cells exposed to the 280 °C treated particles showed an induction of RNA-related processes, a number of mitochondria-associated processes as well as DNA damage response, while the exposure to 580 °C treated HFO particles mainly induced the regulation of intracellular transport. In summary, our analysis based on a highly reproducible automated proteomic sample-preparation procedure shows a diverse cellular response, depending on the soot particle composition. In particular, it was shown that both the molecules of the adsorbate layer as well as particle cores induced strong but different effects in the exposed cells.

Keywords

SILAC Dimethyl labeling Ship diesel exhaust particles Polycyclic aromatic hydrocarbons Oxidative stress Mitochondria 

Abbreviations

DML

Dimethyl labeling

GC

Gas chromatography

GO

Gene ontology

HFO

Heavy fuel oil

HFO280

Heavy fuel oil, 280 °C treatment

HFO580

Heavy fuel oil, 580 °C treatment

LOD

Limit of detection

PAH

Polycyclic aromatic hydrocarbons

PM

Particulate matter

SILAC

Stable isotope labeling by amino acids in cell culture

Notes

Acknowledgments

This work was supported by funds of the Helmholtz virtual institute for complex Molecular Systems in Environmental Health (HICE). We thank Hanns Paur from the KIT, Karlsruhe, Germany for the loan of the CAROLA electrostatic precipitator and Patrick Beaudette as well as Daniel Perez-Hernandez for carefully reading the paper and fruitful discussions. We thank CTC and the Axel Semrau GmbH for the close collaboration in adapting the automation procedures for the CTC-PAL/Chronos setup.

Conflict of Interest

The authors declare no conflict of interest.

Supplementary material

216_2015_8595_MOESM1_ESM.pdf (256 kb)
ESM 1 (PDF 255 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Tamara Kanashova
    • 1
    • 5
  • Oliver Popp
    • 1
  • Jürgen Orasche
    • 2
    • 5
  • Erwin Karg
    • 2
    • 4
  • Horst Harndorf
    • 4
    • 5
  • Benjamin Stengel
    • 4
    • 5
  • Martin Sklorz
    • 2
  • Thorsten Streibel
    • 2
    • 4
  • Ralf Zimmermann
    • 2
    • 3
    • 5
  • Gunnar Dittmar
    • 1
    • 5
  1. 1.Mass spectrometryMax-Delbrück Center for molecular medicineBerlinGermany
  2. 2.Joint Mass Spectrometry CentreUniversity of RostockRostockGermany
  3. 3.Comprehensive Molecular AnalyticsHelmholtz Zentrum München – German Research Center for Environmental Health GmbHOberschleißheimGermany
  4. 4.Chair of Piston Machines and Internal Combustion EnginesUniversity of RostockRostockGermany
  5. 5.HICE – Helmholtz Virtual Institute of Complex Molecular Systems in Environmental HealthNeuherbergGermany

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