Arsenic-induced protein phosphorylation changes in HeLa cells

ALP O., Merino E. J., Caruso J. A.

ANALYTICAL AND BIOANALYTICAL CHEMISTRY, cilt.398, sa.5, ss.2099-2107, 2010 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 398 Sayı: 5
  • Basım Tarihi: 2010
  • Doi Numarası: 10.1007/s00216-010-4128-3
  • Dergi Adı: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.2099-2107
  • Anahtar Kelimeler: Bioanalytical methods, Cell systems/single-cell analysis, Mass spectrometry/ICP-MS, Speciation, HPLC, Genomics/proteomics, ELEMENT MASS-SPECTROMETRY, CAPILLARY LIQUID-CHROMATOGRAPHY, WARFARE AGENTS, BINDING, IDENTIFICATION, SITE, ELECTROPHORESIS, ENRICHMENT, PEPTIDES, TOXICITY
  • Gazi Üniversitesi Adresli: Evet

Özet

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 P-31 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.