A CTAB based method for the preparation of total protein extract of wine spoilage microrganisms for proteomic analysis

Polati, Rita; Zapparoli, Giacomo; Giudici, Paolo; Bossi, Alessandra Maria

doi:10.1016/j.jchromb.2009.02.022

Cited by 11 publications

(11 citation statements)

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“…Subsequently, our group 29 reported a total of 184 spots by analyzing the O. oeni proteoma by pH 3–10 gradient. In this work, we further improved the O. oeni protein detection by using the cationic detergent CTAB 24, which proves to work extremely well and efficiently. In fact, as shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The samples were sonicated in the presence of glass beads (150–212 μm diameter, Sigma) for 5×30 s on ice with 1 min rest in between times and then centrifuged for 10 min at 10 000× g at 4°C. Supernatants were collected and extraction was repeated on pellets as previously described 24. Then nine volumes of double DI water were added to the supernatants and (after centrifugation to remove DNA and polysaccharides) samples were concentrated by Vivaspin 20 (10 kDa MW).…”

Section: Methodsmentioning

confidence: 99%

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Proteomic analysis of Oenococcus oeni freeze‐dried culture to assess the importance of cell acclimation to conduct malolactic fermentation in wine

et al. 2009

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Cultures of Oenococcus oeni, the most important malolactic bacterium, are used to induce malolactic fermentation in wine. Survival assays in two different wines confirmed that cells acclimated for 24 h in half-strength wine-like medium (acclimation medium) enhanced the malolactic performances. To investigate the effect of the pre-incubation phase on cell physiology, a proteomic study was carried out. Total protein extracts of acclimated and non-acclimated cell cultures (control) were analyzed by 2-D-PAGE. A total of 20 out of approximately 400 spots varied significantly. All the spots were identified by MS analysis and most of them were proteins involved in metabolism, transcription/translation processes and stress response. The results revealed the different physiological status between non-acclimated and acclimated cells explaining, in part, their different behavior in wine. Regulation of stress proteins such as heat and cold shock proteins was involved. Moreover, the availability of sugars and amino acids (even if at low concentration) in acclimation medium determined a modulation of energy metabolism enhancing the resistance to stressful conditions (as those that cells find in wine when inoculated). Finally, this proteomic study increased knowledge concerning the physiological changes in freeze-dried culture occurring with pre-inoculation procedures.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Proteomic analysis of Oenococcus oeni freeze‐dried culture to assess the importance of cell acclimation to conduct malolactic fermentation in wine

et al. 2009

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“…We decided to use the diagonal gel electrophoresis method because it was known to give the best data for the high-mass proteome, especially via the cationic detergent cetyl-trimethyl ammonium bromide (CTAB)-PAGE (Braun et al, 2007;Polati et al, 2009;Yamaguchi et al, 2008a, b).…”

Section: Discussionmentioning

confidence: 99%

“…Diagonal poly acrylamide gel electrophoresis method was used for proteins separation, with the anionic SDS and the cationic CTAB detergents- (Braun et al, 2007;Polati et al, 2009;Yamaguchi et al, 2008a, b). A comparison between the HFIP and ACN/TFA0.1% aq extraction procedures was first performed in 1D SDS-PAGE (Fig.…”

Section: Solvent-based Tissue Solubilization Evaluationmentioning

confidence: 99%

HFIP Extraction Followed by 2D CTAB/SDS-PAGE Separation: A New Methodology for Protein Identification from Tissue Sections after MALDI Mass Spectrometry Profiling for Personalized Medicine Research

Longuespée

Tastet

Desmons

et al. 2014

OMICS: A Journal of Integrative Biology

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Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) and profiling technology have become the easiest methods for quickly accessing the protein composition of a tissue area. Unfortunately, the demand for the identification of these proteins remains unmet. To overcome this bottleneck, we combined several strategies to identify the proteins detected via MALDI profiling including on-tissue protein extraction using hexafluoroIsopropanol (1,1,1,3,3,3-hexafluoro-2-propanol, HFIP) coupled with two-dimensional cetyl trimethylammonium bromide/sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D CTAB/SDS-PAGE) for separation followed by trypsin digestion and MALDI-MS analyses for identification. This strategy was compared with an on-tissue bottom-up strategy that we previously developed. The data reflect the complementarity of the approaches. An increase in the number of specific proteins identified has been established. This approach demonstrates the potential of adapted extraction procedures and the combination of parallel identification approaches for personalized medicine applications. The anatomical context provides important insight into identifying biomarkers and may be considered a first step for tissue-based biomarker research, as well as the extemporaneous examination of biopsies during surgery.

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“…Proteins were purified as previously described by Polati et al . [13]. Briefly, nine volumes of double distilled water were added to the supernatants and samples were centrifuged 20,000xg for 10 min to remove DNA and polysaccharides.…”

Section: Methodsmentioning

confidence: 99%

Effects of mushroom and chicory extracts on the shape, physiology and proteome of the cariogenic bacterium Streptococcus mutans

Signoretto

Marchi

Bertoncelli

et al. 2013

BMC Complement Altern Med

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BackgroundDental caries is an infectious disease which results from the acidic demineralisation of the tooth enamel and dentine as a consequence of the dental plaque (a microbial biofilm) accumulation. Research showed that several foods contain some components with antibacterial and antiplaque activity. Previous studies indicated antimicrobial and antiplaque activities in a low-molecular-mass (LMM) fraction of extracts from either an edible mushroom (Lentinus edodes) or from Italian red chicory (Cichorium intybus).MethodsWe have evaluated the antimicrobial mode of action of these fractions on Streptococcus mutans, the etiological agent of human dental caries. The effects on shape, macromolecular syntheses and cell proteome were analysed.ResultsThe best antimicrobial activity has been displayed by the LMM mushroom extract with a bacteriostatic effect. At the MIC of both extracts DNA synthesis was the main macromolecular synthesis inhibited, RNA synthesis was less inhibited than that of DNA and protein synthesis was inhibited only by roughly 50%. The partial inhibition of protein synthesis is compatible with the observed significant increase in cell mass. The increase in these parameters is linked to the morphological alteration with transition from cocci of the untreated control to elongated cells. Interestingly, these modifications were also observed at sub-MIC concentrations. Finally, membrane and cytosol proteome analysis was conducted under LMM mushroom extract treatment in comparison with untreated S. mutans cells. Significant changes were observed for 31 membrane proteins and 20 of the cytosol fractions. The possible role of the changed proteins is discussed.ConclusionsThis report has shown an antibiotic-like mode of action of mushroom and chicory extracts as demonstrated by induced morphogenetic effects and inhibition of specific macromolecular synthesis. This feature as well as the safe use of this extract as result of its natural origin render the LMM both mushroom and chicory extracts suitable for the formulation into products for daily oral hygiene such as mouthwashes or toothpastes.

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A CTAB based method for the preparation of total protein extract of wine spoilage microrganisms for proteomic analysis

Cited by 11 publications

References 23 publications

Proteomic analysis of Oenococcus oeni freeze‐dried culture to assess the importance of cell acclimation to conduct malolactic fermentation in wine

Proteomic analysis of Oenococcus oeni freeze‐dried culture to assess the importance of cell acclimation to conduct malolactic fermentation in wine

HFIP Extraction Followed by 2D CTAB/SDS-PAGE Separation: A New Methodology for Protein Identification from Tissue Sections after MALDI Mass Spectrometry Profiling for Personalized Medicine Research

Effects of mushroom and chicory extracts on the shape, physiology and proteome of the cariogenic bacterium Streptococcus mutans

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