Using the Kinexus integrated platform of proteomics discovery services, the researchers initially analyzed lung lysates that were pooled from similar groups of exposed rats using antibody microarrays that tracked over 500 key cell signaling proteins for their abundance and their phosphorylation status. Phosphorylation acts as an on/off switch for most of the 23,000 proteins encoded by the human genome. Promising leads from these antibody microarrays were confirmed by immunoblotting studies, and they were further monitored using microarrays that were printed with lysates from individual animals as separate spots on the chips. Over 20 signal transduction and stress proteins were found to be consistently and significantly altered with the short term cigarette exposures, and bioinformatics was used to show how these proteins connected within cell signaling networks.
Smoke-altered proteins regulate apoptosis, proliferation, stress response, cell structure, cell migration and inflammation. The female rats demonstrated much higher sensitivity to low doses of cigarette smoke and resultant changes in these protein pathways than their male counterparts. The study utilized proteomic technologies to enable a systematic approach to understanding lung proteins altered by smoke... [PDF] Kinexus Bioinformatics' Press Release -
