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Quantitative proteome profiling of C. burnetii under tetracycline stress conditions.

Vranakis I, De Bock PJ, Papadioti A, Tselentis Y, Gevaert K, Tsiotis G, Psaroulaki A - PLoS ONE (2012)

Bottom Line: Using the MS-driven combined fractional diagonal chromatography proteomics technique, out of the 531 proteins identified, 5 and 19 proteins were found significantly up- and down-regulated respectively, under tetracycline stress.Although the predicted cellular functions of these regulated proteins did not point to known tetracycline resistance mechanisms, our data clearly reveal the plasticity of the proteome of C. burnetii to battle tetracycline stress.Finally, we raise several plausible hypotheses that could further lead to more focused experiments on studying tetracycline resistance in C. burnetii and thus reduced treatment failures of Q fever.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Bacteriology, Parasitology, Zoonoses, and Geographical Medicine, Medical School, University of Crete, Heraklion, Greece. iosif.vranakis@gmail.com

ABSTRACT
The recommended antibiotic regimen against Coxiella burnetii, the etiological agent of Q fever, is based on a semi-synthetic, second-generation tetracycline, doxycycline. Here, we report on the comparison of the proteomes of a C. burnetii reference strain either cultured under control conditions or under tetracycline stress conditions. Using the MS-driven combined fractional diagonal chromatography proteomics technique, out of the 531 proteins identified, 5 and 19 proteins were found significantly up- and down-regulated respectively, under tetracycline stress. Although the predicted cellular functions of these regulated proteins did not point to known tetracycline resistance mechanisms, our data clearly reveal the plasticity of the proteome of C. burnetii to battle tetracycline stress. Finally, we raise several plausible hypotheses that could further lead to more focused experiments on studying tetracycline resistance in C. burnetii and thus reduced treatment failures of Q fever.

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Classification of the 531 identified C. burnetii proteins according to their cellular function.These 531 proteins were identified in both samples; i. e., C. burnetii cultured in presence of tetracycline and C. burnetii cultured with no antibiotic present.
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pone-0033599-g001: Classification of the 531 identified C. burnetii proteins according to their cellular function.These 531 proteins were identified in both samples; i. e., C. burnetii cultured in presence of tetracycline and C. burnetii cultured with no antibiotic present.

Mentions: The COFRADIC technology used here enriches for methionine-containing peptides out of protease-digested proteomes with the aim to reduce the sample complexity before the actual LC-MS/MS analysis, and thereby, attempts to increase the overall proteome coverage [16]. In total, 13,271 MS/MS spectra were identified, 8,208 (62%) of which were linked to peptides containing methionine. These spectra identified 1,998 unique peptides in 800 proteins (Table 1). In order to reduce the number of possible false positive identification, stringent filtering criteria were used on all peptide-to-spectrum matches (Table 2). These filtering criteria reduced the number of identified and quantified proteins to 531, corresponding to 29% of all ORF products (Table S1). These proteins were assigned to 20 functional categories (Figure 1) based on GO terms [17]. Our protein set covers a wide range of cellular functions, which suggests unbiased sampling of bacterial proteins, a requirement for comparative proteomics studies.


Quantitative proteome profiling of C. burnetii under tetracycline stress conditions.

Vranakis I, De Bock PJ, Papadioti A, Tselentis Y, Gevaert K, Tsiotis G, Psaroulaki A - PLoS ONE (2012)

Classification of the 531 identified C. burnetii proteins according to their cellular function.These 531 proteins were identified in both samples; i. e., C. burnetii cultured in presence of tetracycline and C. burnetii cultured with no antibiotic present.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3306420&req=5

pone-0033599-g001: Classification of the 531 identified C. burnetii proteins according to their cellular function.These 531 proteins were identified in both samples; i. e., C. burnetii cultured in presence of tetracycline and C. burnetii cultured with no antibiotic present.
Mentions: The COFRADIC technology used here enriches for methionine-containing peptides out of protease-digested proteomes with the aim to reduce the sample complexity before the actual LC-MS/MS analysis, and thereby, attempts to increase the overall proteome coverage [16]. In total, 13,271 MS/MS spectra were identified, 8,208 (62%) of which were linked to peptides containing methionine. These spectra identified 1,998 unique peptides in 800 proteins (Table 1). In order to reduce the number of possible false positive identification, stringent filtering criteria were used on all peptide-to-spectrum matches (Table 2). These filtering criteria reduced the number of identified and quantified proteins to 531, corresponding to 29% of all ORF products (Table S1). These proteins were assigned to 20 functional categories (Figure 1) based on GO terms [17]. Our protein set covers a wide range of cellular functions, which suggests unbiased sampling of bacterial proteins, a requirement for comparative proteomics studies.

Bottom Line: Using the MS-driven combined fractional diagonal chromatography proteomics technique, out of the 531 proteins identified, 5 and 19 proteins were found significantly up- and down-regulated respectively, under tetracycline stress.Although the predicted cellular functions of these regulated proteins did not point to known tetracycline resistance mechanisms, our data clearly reveal the plasticity of the proteome of C. burnetii to battle tetracycline stress.Finally, we raise several plausible hypotheses that could further lead to more focused experiments on studying tetracycline resistance in C. burnetii and thus reduced treatment failures of Q fever.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Bacteriology, Parasitology, Zoonoses, and Geographical Medicine, Medical School, University of Crete, Heraklion, Greece. iosif.vranakis@gmail.com

ABSTRACT
The recommended antibiotic regimen against Coxiella burnetii, the etiological agent of Q fever, is based on a semi-synthetic, second-generation tetracycline, doxycycline. Here, we report on the comparison of the proteomes of a C. burnetii reference strain either cultured under control conditions or under tetracycline stress conditions. Using the MS-driven combined fractional diagonal chromatography proteomics technique, out of the 531 proteins identified, 5 and 19 proteins were found significantly up- and down-regulated respectively, under tetracycline stress. Although the predicted cellular functions of these regulated proteins did not point to known tetracycline resistance mechanisms, our data clearly reveal the plasticity of the proteome of C. burnetii to battle tetracycline stress. Finally, we raise several plausible hypotheses that could further lead to more focused experiments on studying tetracycline resistance in C. burnetii and thus reduced treatment failures of Q fever.

Show MeSH
Related in: MedlinePlus