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Protein expression profiling of Caco‐2/HT29‐MTX co‐culture after functionalized carbon nanotube exposure
Author(s) -
Lai Xianyin,
Clack James W.,
Ntim Susana Ado,
Mitra Somenath,
Witzmann Frank A.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.863.5
Subject(s) - carbon nanotube , chemistry , caco 2 , tandem mass spectrometry , chromatography , in vitro , biochemistry , mass spectrometry , nanotechnology , materials science
To assess effects of dispersible, functionalized carbon nanotube (CNT) exposure in an in vitro model simulating the digestive tract, Caco‐2/HT29‐MTX cell protein expression was evaluated using liquid chromatography/tandem mass spectrometry (LC‐MS/MS). Three types of CNT, carboxylated single‐wall carbon nanotubes (cSWNT), carboxylated multi‐wall carbon nanotubes (cMWNT), and poly(vinylpyrrolidone) (PVP) polymer functionalized multi‐wall carbon nanotubes (pMWNT), were applied to the co‐cultured cells (75% Caco‐2, 25% HT29‐MTX) at 500 pg/mL and 10 μg/mL for 48 hrs. Proteins were extracted from the cells, reduced, alkylated, and digested. Tryptic peptides were analyzed by LC‐MS/MS. The peptides and proteins were identified using SEQUEST and validated by the Trans‐Proteomic Pipeline. Protein quantification was performed using an in‐house software package, IdentiQuantXL TM . 5007 proteins were identified with >90% confidence and 4199 proteins were quantified. Among these, 803 proteins were differentially expressed (P<0.01). Bioinformatic analysis of the results indicates a significant biological effect of CNTs on Caco‐2/HT29‐MTX protein expression, in the absence of overt toxicity, and merit further study. Supported by NIEHS RC2ES018025.