PMID- 25598225 OWN - NLM STAT- MEDLINE DCOM- 20150413 LR - 20181113 IS - 1879-3185 (Electronic) IS - 0300-483X (Print) IS - 0300-483X (Linking) VI - 329 DP - 2015 Mar 2 TI - Toxicoproteomic analysis of pulmonary carbon nanotube exposure using LC-MS/MS. PG - 80-7 LID - S0300-483X(15)00019-0 [pii] LID - 10.1016/j.tox.2015.01.011 [doi] AB - Toxicoproteomics is a developing field that utilizes global proteomic methodologies to investigate the physiological response as a result of adverse toxicant exposure. The aim of this study was to compare the protein secretion profile in lung bronchoalveolar lavage fluid (BALF) from mice exposed to non-functionalized multi-walled carbon nanotubes (U-MWCNTs) or MWCNTs functionalized by nanoscale Al2O3 coatings (A-MWCNT) formed using atomic layer deposition (ALD). Proteins were identified using liquid chromatography tandem mass spectrometry (LC-MS/MS), and quantified using a combination of two label-free proteomic methods: spectral counting and MS1 peak area analysis. On average 465 protein groups were identified per sample and proteins were first screened using spectral counting and the Fisher's exact test to determine differentially regulated species. Significant proteins by Fisher's exact test (p<0.05) were then verified by integrating the intensity under the extracted ion chromatogram from a single unique peptide for each protein across all runs. A two sample t-test based on integrated peak intensities discovered differences in 27 proteins for control versus U-MWCNT, 13 proteins for control versus A-MWCNT, and 2 proteins for U-MWCNT versus A-MWCNT. Finally, an in-vitro binding experiment was performed yielding 4 common proteins statistically different (p<0.05) for both the in-vitro and in-vivo study. Several of the proteins found to be significantly different between exposed and control groups are known to play a key role in inflammatory and immune response. A comparison between the in-vitro and in-vivo CNT exposure emphasized a true biological response to CNT exposure. CI - Copyright (c) 2015 Elsevier Ireland Ltd. All rights reserved. FAU - Hilton, Gina M AU - Hilton GM AD - Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States. FAU - Taylor, Alexia J AU - Taylor AJ AD - Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States. FAU - McClure, Christina D AU - McClure CD AD - Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, United States. FAU - Parsons, Gregory N AU - Parsons GN AD - Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, United States. FAU - Bonner, James C AU - Bonner JC AD - Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States. FAU - Bereman, Michael S AU - Bereman MS AD - Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States. Electronic address: michaelbereman@ncsu.edu. LA - eng GR - T32 ES007046/ES/NIEHS NIH HHS/United States GR - RC2 ES018772/ES/NIEHS NIH HHS/United States GR - NIEHS R01ES020897/ES/NIEHS NIH HHS/United States GR - NIEHS RC2ES018772/ES/NIEHS NIH HHS/United States GR - R01 ES020897/ES/NIEHS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20150115 PL - Ireland TA - Toxicology JT - Toxicology JID - 0361055 RN - 0 (Acute-Phase Proteins) RN - 0 (Complement C3) RN - 0 (Complement C9) RN - 0 (Histones) RN - 0 (Lipocalin-2) RN - 0 (Lipocalins) RN - 0 (Nanotubes, Carbon) RN - 0 (Oncogene Proteins) RN - 0 (Proteome) RN - 0 (Pulmonary Surfactant-Associated Protein B) RN - 126469-30-5 (Lcn2 protein, mouse) RN - 80295-50-7 (Complement C4b) RN - EC 1.11.1.7 (Peroxidase) RN - EC 3.4.21.- (Lactoferrin) SB - IM MH - Acute-Phase Proteins/genetics/metabolism MH - Animals MH - Bronchoalveolar Lavage Fluid/chemistry MH - Chromatography, Liquid MH - Complement C3/genetics/metabolism MH - Complement C4b/genetics/metabolism MH - Complement C9/genetics/metabolism MH - Histones/genetics/metabolism MH - Lactoferrin/genetics/metabolism MH - Lipocalin-2 MH - Lipocalins/genetics/metabolism MH - Lung/*drug effects/metabolism MH - Mice MH - Mice, Inbred C57BL MH - Nanotubes, Carbon/chemistry/*toxicity MH - Oncogene Proteins/genetics/metabolism MH - Peroxidase/genetics/metabolism MH - Proteome/*metabolism MH - Pulmonary Surfactant-Associated Protein B/genetics/metabolism MH - Tandem Mass Spectrometry MH - Toxicity Tests PMC - PMC4442484 MID - NIHMS690468 OTO - NOTNLM OT - Carbon nanotube OT - Label-free proteomics OT - Pulmonary fibrosis OT - Toxicoproteomics COIS- Conflict of interest The authors declare that there are no conflicts of interest. EDAT- 2015/01/20 06:00 MHDA- 2015/04/14 06:00 PMCR- 2015/05/25 CRDT- 2015/01/20 06:00 PHST- 2014/10/14 00:00 [received] PHST- 2014/12/15 00:00 [revised] PHST- 2015/01/14 00:00 [accepted] PHST- 2015/01/20 06:00 [entrez] PHST- 2015/01/20 06:00 [pubmed] PHST- 2015/04/14 06:00 [medline] PHST- 2015/05/25 00:00 [pmc-release] AID - S0300-483X(15)00019-0 [pii] AID - 10.1016/j.tox.2015.01.011 [doi] PST - ppublish SO - Toxicology. 2015 Mar 2;329:80-7. doi: 10.1016/j.tox.2015.01.011. Epub 2015 Jan 15.