系统生物学技术在生命科学研究中的应用.ppt

Purpose of this study As one of the smallest self-replicating organisms capable of a cell-free existence, M. pneumoniae has been widely used as a model system to evaluate such systems biology approaches as genomics and proteomics Therefore, in the present study, M. pneumoniae infection was also used as a model to examine the applicability of lipidomics in the study of bacterial pathogenesis Results I. Sphingolipid profiles of M. pneumoniae and A549 cells sphingolipids were extracted from M. pneumoniae and A549 cells then subjected to MALDI- TOF-MS analysis molecular ions [M+Na]+, [M+H]+, [M+K]+, or [M+H2O +H]+ for two major classes of sphingolipids, e.g., ceramide (Cer) or sphingomyelin (SM) species corresponding to specific peaks were assigned Representative mass spectra of sphingolipids in M. pneumoniae and A549 cells. Characteristic ions (m/z) M. pneumoniae Uninfected A549 cells 502.9 Cer[d18:1C12:1+Na]+ + + 506.9 Cer[d18:1C11:0+K]+ + + 507.8 Cer[d16:1C16:1+H]+ + - 508.9 Cer[d18:1C14:1+H]+ - + 511.1 Cer[d18:1C14:0 +H]+ + - 518.8 Cer[d18:1C12:1+K]+ - + 528.8 Cer[d18:1C14:0+H2O+H]+ + + 539.0 Cer[d18:1C16:0 +H]+ + + 549.7 Cer[d18:1C17:1+H]+ + - 574.5 Cer[d18:1C16:1+K]+ + - 701.6 SM[d18:1C16:1+H]+ + + 723.5 SM[d18:1C16:1+Na]+ + + 729.6 SM[d18:1C18:1+H]+ + - 751.5 SM[d18:1C18:1+Na]+ + - 811.4 SM[d18:1C22:0+Na]+ + + 833.3 SM[d18:0C24h:0+H]+ + - Sphingolipids species in M. pneumoniae and uninfected A549 cells M. pneumoniae and A549 cells share many common sphingolipid molecules yet 7 peaks were only present in M. pneumoniae they were defined as a marker of M. pneumoniae sphingolipids On the other hand, several peaks were only found in A549 cells II. M. pneumoniae infection induces significant changes in cellular sphingolipid composition M. pneumoniae at concentrations of 1 , 10 , and 100 CFU/cell Sphingolipids were then extracted from cells at 2 ,12 , and 24 h post infection, and analyzed by MALDI-TOF-MS Representative mass spectra of sphingolipids in infected and un