Proteomic Investigation of the Time Course Responses of RAW 264.7 Macrophages to Infection with Salmonella enterica

Journal Article · Sat Aug 01 04:00:00 EDT 2009 · Infection and Immunity, 77(8):3227-3233

DOI:https://doi.org/10.1128/IAI.00063-09· OSTI ID:964228

Shi, Liang; Chowdhury, Saiful M; Smallwood, Heather S; Yoon, Hyunjin; Mottaz-Brewer, Heather M; Norbeck, Angela D; McDermott, Jason E; Clauss, Therese RW; Heffron, Fred; Smith, Richard D; Adkins, Joshua N

Macrophages plan important roles in controlling Salmonella-mediated systemic infection. To investigate the responses of macrophages to Salmonella infection, we infected RAW 264.7 macrophages with Salmonella enterica serovar Typhimurium (STM) and then performed a comparative liquid chromatography-tandem mass spectrometry [LC-MS(/MS)]-based proteomics analysis of the infected macrophages. A total of 1006 macrophage and 115 STM proteins were indentified from this study. Most of STM proteins were found at late stage of the time course of infection, consistent with the fact that STM proliferates inside RAW 264.7 macrophages. Majority of the identified macrophage proteins were house keeping-related, including cytoplasmic superoxide dismutase 1 (SOD1), whose peptide abundances were relatively constant during the time course of infection. Compared to those in no infection control, the peptide abundances of 244 macrophage proteins (or 24% of total indentified macrophage proteins) changed considerably after STM infection. The functions of these STM infection-affected macrophage proteins were diverse and ranged from production of antibacterial nitric oxide (i.e., inducible nitric oxide synthase or iNOS) or production of prostaglandin H2 (i.e., prostaglandin-endoperoxide synthase 2, also know as cyclooxygenase-2 or COX-2) to regulation of intracellular traffic (e.g., sorting nexin or SNX 5, 6 and 9), demonstrating a global impact of STM infection on macrophage proteome. Western-blot analysis not only confirmed the LC-MS(/MS) results of SOD1, COX-2 and iNOS, but also revealed that the protein abundances of mitochondrial SOD2 increased after STM infection, indicating an infection-induced oxidative stress in mitochondria.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 964228
• Report Number(s): PNNL-SA-62432; 11491
• Journal Information: Infection and Immunity, 77(8):3227-3233, Journal Name: Infection and Immunity, 77(8):3227-3233 Journal Issue: 8 Vol. 77; ISSN 0019-9567; ISSN INFIBR
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Environmental Molecular Sciences Laboratory
MACROPHAGES
MASS SPECTROSCOPY
MITOCHONDRIA
NITRIC OXIDE
PEPTIDES
PRODUCTION
PROSTAGLANDINS
PROTEINS
Proteomics
host-pathogen interaction
REGULATIONS
SALMONELLA
SORTING
SUPEROXIDE DISMUTASE