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Mass spectrometry-based proteomic approaches to study pathogenic bacteria-host interactions.

Yang Y, Hu M, Yu K, Zeng X, Liu X - Protein Cell (2015)

Bottom Line: Within the last decade, mass spectrometry (MS)-based proteomics has become a powerful and effective approach to better understand complex and dynamic host-pathogen interactions at the protein level.Herein we will review the recent progress in proteomic analyses towards bacterial infection of their mammalian host with a particular focus on enteric pathogens.Large-scale studies of dynamic proteomic alterations during infection will be discussed from the perspective of both pathogenic bacteria and host cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Analytical Chemistry and Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.

ABSTRACT
Elucidation of molecular mechanisms underlying host-pathogen interactions is important for control and treatment of infectious diseases worldwide. Within the last decade, mass spectrometry (MS)-based proteomics has become a powerful and effective approach to better understand complex and dynamic host-pathogen interactions at the protein level. Herein we will review the recent progress in proteomic analyses towards bacterial infection of their mammalian host with a particular focus on enteric pathogens. Large-scale studies of dynamic proteomic alterations during infection will be discussed from the perspective of both pathogenic bacteria and host cells.

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Related in: MedlinePlus

A general workflow of proteomic studies of host-pathogen interactions. Upon infection of host cells (with intracellular bacteria as an example), bacterial pathogens were physically isolated from host cells and subjected to high-throughput proteomic analyses by LC-MS-based approaches. Host cellular components can also be analyzed, though extensive sample fractionations were necessary for sufficient proteome coverage (i.e., subcellular enrichment and protein gel separation). Protein samples are enzymatically digested into peptide mixtures prior to LC-MS measurements for both qualitative and quantitative analyses
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Fig1: A general workflow of proteomic studies of host-pathogen interactions. Upon infection of host cells (with intracellular bacteria as an example), bacterial pathogens were physically isolated from host cells and subjected to high-throughput proteomic analyses by LC-MS-based approaches. Host cellular components can also be analyzed, though extensive sample fractionations were necessary for sufficient proteome coverage (i.e., subcellular enrichment and protein gel separation). Protein samples are enzymatically digested into peptide mixtures prior to LC-MS measurements for both qualitative and quantitative analyses

Mentions: Over the years, two-dimensional (2-D) gel electrophoresis followed by MS identifications of individual protein spots has been utilized for the prototype proteomic studies (Curreem et al., 2012; Rabilloud et al., 2010). In addition to being time-consuming, however, 2-D gel can never meet the challenges of complex biological samples due to its rather limited sensitivity and dynamic range (Jafari et al., 2012). Fortunately within the last decade, MS-based proteomics has evolved into a high-sensitivity high-throughput approach for quantitative examination of proteins from any biological system on a global scale (Fig. 1). The core platform of this technology is liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), in which complex protein samples are enzymatically digested into peptides prior to chromatographic separation and MS identifications (Cravatt et al., 2007). Importantly, the field of MS-based proteomics has witnessed major technical breakthroughs such as the advent of high-resolution Orbitrap mass spectrometers. Currently, the state-of-the-art proteomic technologies are capable of measuring several thousands of proteins within a few hours, which permits us to significantly increase the analytical depth as well as coverage in complex proteome analyses (Ding et al., 2013; Geiger et al., 2012; Weekes et al., 2014).Figure 1


Mass spectrometry-based proteomic approaches to study pathogenic bacteria-host interactions.

Yang Y, Hu M, Yu K, Zeng X, Liu X - Protein Cell (2015)

A general workflow of proteomic studies of host-pathogen interactions. Upon infection of host cells (with intracellular bacteria as an example), bacterial pathogens were physically isolated from host cells and subjected to high-throughput proteomic analyses by LC-MS-based approaches. Host cellular components can also be analyzed, though extensive sample fractionations were necessary for sufficient proteome coverage (i.e., subcellular enrichment and protein gel separation). Protein samples are enzymatically digested into peptide mixtures prior to LC-MS measurements for both qualitative and quantitative analyses
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4383758&req=5

Fig1: A general workflow of proteomic studies of host-pathogen interactions. Upon infection of host cells (with intracellular bacteria as an example), bacterial pathogens were physically isolated from host cells and subjected to high-throughput proteomic analyses by LC-MS-based approaches. Host cellular components can also be analyzed, though extensive sample fractionations were necessary for sufficient proteome coverage (i.e., subcellular enrichment and protein gel separation). Protein samples are enzymatically digested into peptide mixtures prior to LC-MS measurements for both qualitative and quantitative analyses
Mentions: Over the years, two-dimensional (2-D) gel electrophoresis followed by MS identifications of individual protein spots has been utilized for the prototype proteomic studies (Curreem et al., 2012; Rabilloud et al., 2010). In addition to being time-consuming, however, 2-D gel can never meet the challenges of complex biological samples due to its rather limited sensitivity and dynamic range (Jafari et al., 2012). Fortunately within the last decade, MS-based proteomics has evolved into a high-sensitivity high-throughput approach for quantitative examination of proteins from any biological system on a global scale (Fig. 1). The core platform of this technology is liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), in which complex protein samples are enzymatically digested into peptides prior to chromatographic separation and MS identifications (Cravatt et al., 2007). Importantly, the field of MS-based proteomics has witnessed major technical breakthroughs such as the advent of high-resolution Orbitrap mass spectrometers. Currently, the state-of-the-art proteomic technologies are capable of measuring several thousands of proteins within a few hours, which permits us to significantly increase the analytical depth as well as coverage in complex proteome analyses (Ding et al., 2013; Geiger et al., 2012; Weekes et al., 2014).Figure 1

Bottom Line: Within the last decade, mass spectrometry (MS)-based proteomics has become a powerful and effective approach to better understand complex and dynamic host-pathogen interactions at the protein level.Herein we will review the recent progress in proteomic analyses towards bacterial infection of their mammalian host with a particular focus on enteric pathogens.Large-scale studies of dynamic proteomic alterations during infection will be discussed from the perspective of both pathogenic bacteria and host cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Analytical Chemistry and Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.

ABSTRACT
Elucidation of molecular mechanisms underlying host-pathogen interactions is important for control and treatment of infectious diseases worldwide. Within the last decade, mass spectrometry (MS)-based proteomics has become a powerful and effective approach to better understand complex and dynamic host-pathogen interactions at the protein level. Herein we will review the recent progress in proteomic analyses towards bacterial infection of their mammalian host with a particular focus on enteric pathogens. Large-scale studies of dynamic proteomic alterations during infection will be discussed from the perspective of both pathogenic bacteria and host cells.

Show MeSH
Related in: MedlinePlus