Differential mitochondrial proteomic analysis of A549 cells infected with avian influenza virus subtypes H5 and H9

Abstract Background Both the highly pathogenic avian influenza (HPAI) H5N1 and low (LPAI) H9N2 viruses have been reported to cross species barriers infect humans. can cause severe damage are associated with a high mortality rate, but do not such outcomes. Our purpose was use proteomics technology study differential expression of mitochondrial-related proteins related virus infections. Methods According determined viral infection titer, A549 cells were infected 1 multiplicity virus, mitochondria extracted after 24 h incubation. The protein from lysed analyzed by BCA method determine concentration, as well SDS-PAGE (preliminary analysis), two-dimensional gel electrophoresis, mass spectrometry. Differential spots selected, Western blotting performed verify results. identified subjected GO analysis for subcellular localization, KEGG functional classification signaling pathways assessment, STRING association network construction. Results In 2-D electrophoresis analysis, 227 detected in H5N1-infected group, 169 H9N2-infected group. Protein further spectrometry identification removal redundancy, 32 differentially expressed identified. Compared group had 16 upregulated mitochondrial downregulated proteins. 70-kDa heat shock analogs, short-chain enoyl-CoA hydratase, malate dehydrogenase, ATP synthase verified blot, results consistent findings. Functional indicated that these primarily involved apoptosis metabolism. Conclusions their eight led host T cell activation, antigen presentation, stress response, synthesis reduction, leading higher pathogenicity than H9N2.