Identification of in vitro differential cell secretions due to cigarette smoke condensate exposure using nanoflow capillary liquid chromatography and high-resolution mass spectrometry

Secreted proteins, the secretome, can be isolated from biological fluids (e.g., blood) and are often responsible for the regulation of biological processes such as cell signaling, growth, and apoptosis. The identification of secreted proteins can lead to an understanding of disease mechanisms and th...

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Bibliographic Details
Published in:	Analytical and bioanalytical chemistry Vol. 391; no. 8; pp. 2845 - 2856
Main Authors:	Flora, Jason W, Edmiston, Jeff, Secrist, Rebecca, Li, Guoya, Rana, Gaurav S. J. B, Langston, Timothy B, McKinney, Willie
Format:	Journal Article
Language:	English
Published:	Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01-08-2008 Springer-Verlag
Subjects:	Amino Acid Sequence Analytical Chemistry Biochemistry Bodily Secretions - chemistry Cell Survival Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science Chromatography, Liquid - instrumentation Chromatography, Liquid - methods Cigarette smoke total particulate matter endothelial cells Endothelial Cells - metabolism Food Science Fourier transform ion cyclotron resonance mass spectrometry Humans Inhalation Exposure Laboratory Medicine LTQ FT mass spectrometer Mass Spectrometry - instrumentation Mass Spectrometry - methods Molecular Sequence Data Monitoring/Environmental Analysis Nanoflow capillary liquid chromatography Nanotechnology Nicotiana Original Paper Peptides - chemistry Proteomics - instrumentation Proteomics - methods Secreted proteins Secretome Smoke - adverse effects Cigarette smoke total particulate matter LTQ FT mass spectrometer Secretome Fourier transform ion cyclotron resonance mass spectrometry Nanoflow capillary liquid chromatography Secreted proteins Endothelial cells
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Summary:	Secreted proteins, the secretome, can be isolated from biological fluids (e.g., blood) and are often responsible for the regulation of biological processes such as cell signaling, growth, and apoptosis. The identification of secreted proteins can lead to an understanding of disease mechanisms and they can serve as early candidate biomarkers of disease and exposure. However, it is time-consuming and costly to conduct in vivo interrogations of the human secretome. The purpose of this article is to provide a detailed description of a rapid in vitro technique for the analysis of differential protein secretion due to exposure to smoking-machine-generated cigarette smoke (CS) condensate (total particulate matter, TPM). Endothelial cells were exposed to CS-TPM, the supernatant was collected, and the secretome was elucidated by nano liquid chromatography coupled with high-resolution mass spectrometry. A total of 1,677 unique peptides were identified in the cell culture supernatants. Several proteins were differentially expressed following CS-TPM exposure that relate to several biological processes, such as metabolism, development, communication, response to stimulus, and response to stress.
Bibliography:	http://dx.doi.org/10.1007/s00216-008-2197-3 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	1618-2642 1618-2650
DOI:	10.1007/s00216-008-2197-3