title: Toxicoproteomic analysis of pulmonary carbon nanotube exposure using LC-MS/MS.
abstract: 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.
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States.
authors.forename: Gina M
authors.initials: GM
authors.lastname: Hilton
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States.
authors.forename: Alexia J
authors.initials: AJ
authors.lastname: Taylor
authors.affiliation: Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, United States.
authors.forename: Christina D
authors.initials: CD
authors.lastname: McClure
authors.affiliation: Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, United States.
authors.forename: Gregory N
authors.initials: GN
authors.lastname: Parsons
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States.
authors.forename: James C
authors.initials: JC
authors.lastname: Bonner
authors.affiliation: Department of Biological Sciences, Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC, United States. Electronic address: michaelbereman@ncsu.edu.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Bereman
mesh_terms: D000209:Acute-Phase Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D002853:Chromatography, Liquid; D003176:Complement C3 / Q000235:genetics / Q000378:metabolism; D015935:Complement C4b / Q000235:genetics / Q000378:metabolism; D003186:Complement C9 / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D007781:Lactoferrin / Q000235:genetics / Q000378:metabolism; D000071068:Lipocalin-2; D054834:Lipocalins / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D037742:Nanotubes, Carbon / Q000737:chemistry / Q000633:toxicity*; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D009195:Peroxidase / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D037701:Pulmonary Surfactant-Associated Protein B / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D018675:Toxicity Tests
chemical_list: D000209:Acute-Phase Proteins; D003176:Complement C3; D003186:Complement C9; D006657:Histones; D000071068:Lipocalin-2; D054834:Lipocalins; D037742:Nanotubes, Carbon; D015513:Oncogene Proteins; D020543:Proteome; D037701:Pulmonary Surfactant-Associated Protein B; C071332:Lcn2 protein, mouse; D015935:Complement C4b; D009195:Peroxidase; D007781:Lactoferrin
keywords: Carbon nanotube; Label-free proteomics; Pulmonary fibrosis; Toxicoproteomics