Skip to main content

Advertisement

Log in

Arsenic-induced protein phosphorylation changes in HeLa cells

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Arsenic is well documented as a chemotherapeutic agent capable of inducing cell death while at the same time is considered a human carcinogen and an environmental contaminant. Although arsenic toxicity is well known and has formed an impressive literature over the time, little is known about how its effects are exerted at the proteome level. Protein phosphorylation is an important post-translational modification involved in the regulation of cell signaling and likely is altered by arsenic treatment. Despite the importance of phosphorylation for many regulatory processes in cells, the identification and characterization of phosphorylation, as effected by arsenic through mass spectrometric detection, are not fully studied. Here, we identify phosphorylated proteins, which are related to post-translational modifications after phenylarsine oxide (PAO) inoculation to HeLa cells. PAO was chosen because of its high cytotoxicity, measured earlier in these labs. In this study, size exclusion chromatography coupled to inductively coupled plasma mass spectrometry (SEC-ICP-MS) is used to establish several molecular weight fractions with phosphorylated proteins by monitoring 31P signal vs. time via ICP-MS. SEC-ICP-MS fractions are collected and then separated by the nano-LC-CHIP/ITMS system for peptide determination. Spectrum Mill and MASCOT protein database search engines are used for protein identification. Several phosphorylation sites and proteins related to post-translational modifications are also identified.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Hirano S, Cui X, Li S, Kanno S, Kobayashi Y, Hayakawa T, Shraim A (2003) Arch Toxicol 77:305–312

    CAS  Google Scholar 

  2. Leermakers M, Baeyens W, De Gieter M, Smedts B, Meert C, De Bisschop HC, Morabito R, Quevauviller P (2006) TrAC, Trends Anal Chem 25:1–10

    Article  CAS  Google Scholar 

  3. Tornes JA, Opstad AM, Johnsen BA (2006) Sci Total Environ 356:235–246

    Article  CAS  Google Scholar 

  4. Stock T (1996) Sea-dumped chemical weapons and the chemical weapons convention. In: Kafka AV (ed) Sea-dumped chemical weapons: aspects, problems and solutions. Kluwer Academic, Dordrecht

    Google Scholar 

  5. Kroening KK, Solivio MJV, Garcia-Lopez G, Puga A, Caruso JA (2008) Metallomics 1:59–66

    Article  Google Scholar 

  6. Pitten FA, Muller G, Konig P, Schmidt D, Thurow K, Kramer A (1999) Sci Total Environ 226:237–245

    Article  CAS  Google Scholar 

  7. Charoensuk V, Gati WP, Weinfeld M, Le XC (2009) Toxicol Appl Pharmacol 239:64–70

    Article  CAS  Google Scholar 

  8. Hirano S, Kobayashi Y, Hayakawa T, Cui X, Yamamoto M, Kanno S, Shraim A (2005) Arch Toxicol 79:54–61

    Article  CAS  Google Scholar 

  9. Sahara N, Takeshita A, Kobayashi M, Shigeno K, Nakamura S, Shinjo K, Naito K, Maekawa M, Horii T, Ohnishi K, Kitamura K, Naoe T, Hayashi H, Ohno R (2004) Leuk Lymphoma 45:987–995

    Article  CAS  Google Scholar 

  10. Cline DJ, Thorpe C, Schneider JP (2003) J Am Chem Soc 125:2923–2929

    Article  CAS  Google Scholar 

  11. Wang Z, Zhang H, Li XF, Le XC (2007) Rapid Commun Mass Spectrom 21:3658–3666

    Article  CAS  Google Scholar 

  12. Kitchin KT, Wallace K (2005) Toxicol Appl Pharmacol 206:66–72

    Article  CAS  Google Scholar 

  13. Guo Y, Ling Y, Thomson BA, Siu KWM (2005) J Am Soc Mass Spectrom 16:1787–1794

    Article  CAS  Google Scholar 

  14. Ngu TT, Stillman MJ (2006) J Am Chem Soc 128:12473–12483

    Article  CAS  Google Scholar 

  15. Pinkse MWH, Uitto PM, Hilhorst MJ, Ooms B, Heck AJR (2004) Anal Chem 76:3935–3943

    Article  CAS  Google Scholar 

  16. Wolschin F, Wienkoop S, Weckwerth W (2005) Proteomics 5:4389–4397

    Article  CAS  Google Scholar 

  17. Loyet KM, Stults JT, Arnott D (2005) Mol Cell Proteomics 4:235–245

    Article  CAS  Google Scholar 

  18. Schlosser A, Vanselow JT, Kramer A (2005) Anal Chem 77:5243–5250

    Article  CAS  Google Scholar 

  19. Chalmers MJ, Kolch W, Emmett MR, Marshall AG, Mischak H (2004) J Chromatogr B Anal Technol Biomed Life Sci 803:111–120

    Article  CAS  Google Scholar 

  20. Zeller M, Konig S (2004) Anal Bioanal Chem 378:898–909

    Article  CAS  Google Scholar 

  21. Wind M, Edler M, Jakubowski N, Linscheid M, Wesch H, Lehmann WD (2001) Anal Chem 73:29–35

    Article  CAS  Google Scholar 

  22. Marshall P, Heudi O, Bains S, Freeman HN, Abou-Shakra F, Reardon K (2002) Analyst 127:459–461

    Article  CAS  Google Scholar 

  23. Wind M, Feldmann I, Jakubowski N, Lehmann WD (2003) Electrophoresis 24:1276–1280

    Article  CAS  Google Scholar 

  24. Wind M, Wesch H, Lehmann WD (2001) Anal Chem 73:3006–3010

    Article  CAS  Google Scholar 

  25. Wind M, Gosenca D, Kubler D, Lehmann WD (2003) Anal Biochem 317:26–33

    Article  CAS  Google Scholar 

  26. Wind M, Kelm O, Nigg EA, Lehmann WD (2002) Proteomics 2:1516–1523

    Article  CAS  Google Scholar 

  27. Abdullaev FI, Rivera-Luna R, García-Carrancá A, Ayala-Fierro F, Espinosa-Aguirre JJ (2001) Mutat Res Genet Toxicol Environ Mutagen 493:31–38

    Article  CAS  Google Scholar 

  28. Chen XW, Zou AM, Chen ML, Wang JH, Dasgupta PK (2009) Anal Chem 81:1291–1296

    Article  CAS  Google Scholar 

  29. Gerlach VL, Feaver WJ, Fischhaber PL, Friedberg EC (2001) J Biol Chem 276:92–98

    Article  CAS  Google Scholar 

  30. Briggs SD, Bryant SS, Jove R, Sanderson SD, Smithgall TE (1995) J Biol Chem 270:14718–14724

    Article  CAS  Google Scholar 

  31. Tourrière H, Chebli K, Zekri L, Courselaud B, Blanchard JM, Bertrand E, Tazi J (2003) J Cell Biol 160:823–831

    Article  Google Scholar 

  32. Moon SY, Zheng Y (2003) Trends Cell Biol 13:13–22

    Article  CAS  Google Scholar 

  33. Beausoleil SA, Jedrychowski M, Schwartz D, Elias JE, Villén J, Li J, Cohn MA, Cantley LC, Gygi SP (2004) Proc Natl Acad Sci U S A 101:12130–12135

    Article  CAS  Google Scholar 

  34. Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M (2006) Cell 127:635–648

    Article  CAS  Google Scholar 

  35. Shen X, Valencia CA, Szostak J, Dong B, Liu R (2005) Proc Natl Acad Sci U S A 102:5969–5974

    Article  CAS  Google Scholar 

  36. Cosen-Binker LI, Kapus A (2006) Physiology 21:352–361

    Article  CAS  Google Scholar 

  37. Liu H, Sugiura M, Nava VE, Edsall LC, Kono K, Poulton S, Milstien S, Kohama T, Spiegel S (2000) J Biol Chem 275:19513–19520

    Article  CAS  Google Scholar 

  38. Conus NM, Hannan KM, Cristiano BE, Hemmings BA, Pearson RB (2002) J Biol Chem 277:38021–38028

    Article  CAS  Google Scholar 

  39. Ostergaard E (2008) J Inherit Metab Dis 31:226–229

    Article  CAS  Google Scholar 

  40. Cvetkovic A, Angeli Lal Menon A, Thorgersen M, Scott J, Poole F II, Jenney F Jr, Lancaster W, Praissman J, Shanmukh S, Vaccaro B, Trauger S, Kalisiak E, Apon J, Siuzdak G, Yannone S, Tainer J, Adams M (2010) Nature. doi:10.1038/nature09265

    Google Scholar 

Download references

Acknowledgements

The authors are grateful to Agilent Technologies for their continuing support via chromatographic and mass spectrometric instrumentation. OA is particularly thankful to the Scientific and Technical Research Council of Turkey (TUBITAK) for post-doctoral fellowship support.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Joseph A. Caruso.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Alp, O., Merino, E.J. & Caruso, J.A. Arsenic-induced protein phosphorylation changes in HeLa cells. Anal Bioanal Chem 398, 2099–2107 (2010). https://doi.org/10.1007/s00216-010-4128-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-010-4128-3

Keywords

Navigation