Project description:An opportunistic intracellular pathogen Mycobacterium intracellulare, a member of the nontuberculous mycobacteria (NTM) cluster, causes respiratory disease in immunosuppressed hosts, including companion dogs. The purpose of this study is to investigate a host-M.intracellulare interactome in canine monocyte-derived macrophages during the infection.
Project description:Detection of species-specific proteotypic peptides for accurate and easy characterization of infectious non-tuberculous mycobacteria such as Mycobacterium intracellulare is essential. Therefore, we carried out an in-depth global proteomic experiment using M. intracellulare ATCC 13950 strain followed by proteome database search and spectral library generation. The lysate was subjected to in-solution proteomic sample preparation and fractionated using an off-line C18 StageTip. Each fraction was acquired in technical triplicates using a 180 min data-dependent acquisition (DDA) method in Orbitrap Fusion Tribrid (Thermo Scientific) mass spectrometer. The resulting raw DDA data were searched against the M. intracellulare proteome database using Proteome Discoverer and FragPipe. The resulting peptide spectrum matches were converted into a spectral library using BiblioSpec.
