LXQ分析杜克嗜血杆菌细胞裂解液中的蛋白质

简介：　Determine the feasibility of analyzing a complex mixture, represented by a whole cell lysate of H. ducreyi, on the LXQ linear ion trap using a simple, 1D-SDS PAGE desalting and protein fractionation procedure. Study the effect of non-MS parameters, such as sample complexity and chromatographic resolution on the number and quality of the spectra generated and the resulting proteins identified, with particular emphasis on membrane and high molecular weight proteins.

仪器：LXQ

结论：This study was performed to demonstrate a rapid and sensitive protocol for the analysis of a whole cell lysate using a quick and simple 1D-SDS PAGE fractionation step followed by LC/MSn analysis using the LXQ linear ion trap mass spectrometer. The data clearly shows both the speed and sensitivity of the LXQ, ideal for analyzing complex proteomics samples. Lengthening the chromatographic gradient as well as reducing the complexity of thesample via gel fractionation increased the number of SEQUEST-searchable MSn spectra and subsequent proteins identified.

In this experiment, seventy-nine proteins were identified in a 45-min run, 222 proteins were identified by extending the gradient to 150 min, and 360 proteins were found in total by dividing the whole cell lysate gel band into five distinct slices and using the 150-min gradient.A variety of small and large membrane proteins, which are linked to pathogenesis, and large molecular weight proteins, which are often lost using 2D-gels, were identified. By employing the longer gradients, two very large supernatant proteins (>450 kDa) were also identified.