Targeting of Bacillus anthracis interaction factors for human macrophages using two-dimensional gel electrophoresis

Seo, Gwi Moon; Kim, Seong Joo; Kim, Ji Chon; Nam, Deok Hwa; Yoon, Mi Young; Koo, Bon Sung; Chai, Young Gyu

doi:10.1016/j.bbrc.2004.07.190

Cited by 9 publications

(5 citation statements)

References 16 publications

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“…To analyze cellular proteins, proteomic techniques were performed as previously described [9]. Isoelectric focusing (IEF) was performed using immobilized pH gradient (IPG) dry strips (4-10 NL IPG, 24 cm; Genomine, Korea) that were rehydrated for 16 h with 1.2 mg of sample at 20 o C. After equilibration of the IPG strips, gel electrophoresis was conducted on sodium dodecyl sulfate (SDS) polyacrylamide gels (20 × 24 cm, 10-16%), and the gels were stained with Coomassie brilliant blue [22,23].…”

Section: De Gel Analysis and Spot Quantitationmentioning

confidence: 99%

Proteomic Analysis of the Oxidative Stress Response Induced by Low-Dose Hydrogen Peroxide in Bacillus anthracis

Kim

Kim²,

Jung³

et al. 2013

J. Microbiol. Biotechnol.

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Anthrax is a bacterial disease caused by the aerobic sporeforming bacterium Bacillus anthracis, which is an important pathogen owing to its ability to be used as a terror agent. B. anthracis spores can escape phagocytosis and initiate the germination process even in antimicrobial conditions, such as oxidative stress. To analyze the oxidative stress response in B. anthracis and thereby learn how to prevent antimicrobial resistance, we performed protein expression profiling of B. anthracis strain HY1 treated with 0.3 mM hydrogen peroxide using a comparative proteomics-based approach. The results showed a total of 60 differentially expressed proteins; among them, 17 showed differences in expression over time. We observed time-dependent changes in the production of metabolic and repair/protection signaling proteins. These results will be useful for uncovering the metabolic pathways and protection mechanisms of the oxidative response in B. anthracis.

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Section: De Gel Analysis and Spot Quantitationmentioning

confidence: 99%

Proteomic Analysis of the Oxidative Stress Response Induced by Low-Dose Hydrogen Peroxide in Bacillus anthracis

Kim

Kim²,

Jung³

et al. 2013

J. Microbiol. Biotechnol.

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“…A noncapsulated, toxin‐producing B. anthracis Sterne (pXO1 + pXO2 − ) and delta Sterne (pXO1 − pXO2 − ) were used for this study (Seo et al. 2004).…”

Section: Methodsmentioning

confidence: 99%

Comparative analysis of virulence factors secreted by Bacillus anthracis Sterne at host body temperature

Kim

Shahid

Park

et al. 2012

Letters in Applied Microbiology

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Aims: For the analysis of virulence factors produced and secreted by Bacillus anthracis vegetative cells during mammalian host infection, we evaluated the secretome of B. anthracis Sterne exposed to host‐specific factors specifically to host body temperature. Methods and Results: We employed a comparative proteomics‐based approach to analyse the proteins secreted by B. anthracis Sterne under host‐specific body temperature conditions. A total of 17 proteins encoded on a single chromosome and the pXO1 plasmid were identified by peptide mass fingerprinting. Multiple algorithms were used to predict the secretion mechanisms of the detected proteins in B. anthracis. Conclusions: Several putative virulence factors and known factors responsible for sporulation were differentially regulated, including CodY, pXO1‐130 and BA1952, revealing insights into temperature cues in the B. anthracis secretome. Significance and Impact of the Study: This study identified temperature‐regulated proteins. Further studies aimed at understanding the physical and functional roles of these proteins in infection and control by elevated temperatures will contribute to detection, diagnostics and prophylaxis.

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“…Selected protein spots were digested enzymatically in-gel in a manner similar to that described by Seo et al (2004Seo et al ( , 2008 using modified porcine trypsin (Promega, USA). Gel pieces were washed with 50% acetonitrile to remove sodium dodecyl sulfate (SDS) and salt and dried to remove the solvent.…”

Section: In-gel Digestion Maldi-tof Ms Analysis and Database Searchmentioning

confidence: 99%

Comparative proteome analysis of Bacillus anthracis with pXO1 plasmid content

et al. 2010

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Bacillus anthracis the causative agent of anthrax, is an important pathogen among the Bacillus cereus group of species because of its physiological characteristics and its importance as a biological warfare agent. Tripartite anthrax toxin proteins and a poly-D-glutamic acid capsule are produced by B. anthracis vegetative cells during mammalian hosts infection and when cultured in conditions that are thought to mimic the host environment. To identify the factors regulating virulence in B. anthracis the whole cell proteins were extracted from two B. anthracis strains and separated by narrow range immobilized pH gradient (IPG) strips (pH 4-7), followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins that were differentially expressed were identified by the peptide fingerprinting using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). A total of 23 proteins were identified as being either upregulated or downregulated in the presence or absence of the virulence plasmid pXO1. Two plasmid encoded proteins and 12 cellular proteins were identified and documented as potential virulence factors.

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Targeting of Bacillus anthracis interaction factors for human macrophages using two-dimensional gel electrophoresis

Cited by 9 publications

References 16 publications

Proteomic Analysis of the Oxidative Stress Response Induced by Low-Dose Hydrogen Peroxide in Bacillus anthracis

Proteomic Analysis of the Oxidative Stress Response Induced by Low-Dose Hydrogen Peroxide in Bacillus anthracis

Comparative analysis of virulence factors secreted by Bacillus anthracis Sterne at host body temperature

Comparative proteome analysis of Bacillus anthracis with pXO1 plasmid content

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