Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH)

Kinoshita, Hideki; Imoto, Shun; Suda, Yoshihito; Ishida, Masaharu; Watanabe, Masamichi; Kawai, Yasushi; Kitazawa, Haruki; Miura, Koh; Horii, Akira; Saito, Tadao

doi:10.1111/j.1740-0929.2012.01054.x

Cited by 15 publications

(10 citation statements)

References 45 publications

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“…A 4Á18-fold increased abundance of glyceraldehyde 3-phosphate dehydrogenase (Gap) was observed in strain DRR12 in response to acid (Table 3). Association of Gap to the cell envelope has been previously reported in other bacteria (Pancholi and Fischetti 1992;Ruiz et al 2009) including lactobacilli (Antikainen et al 2007;Nezhad et al 2012;Kinoshita et al 2013). Interestingly, Antikainen et al (2007) observed that Gap is associated to the cell envelope in Lactobacillus crispatus at acidic pH, whereas at neutral pH is rapidly released to the media and that Gap interacts with negatively charged LTAs.…”

Section: Discussionmentioning

confidence: 65%

Characterization of the response to low pH ofLactobacillus caseiΔRR12, a mutant strain with low D-alanylation activity and sensitivity to low pH

et al. 2014

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Aims To identify the differences that account for the acid sensitivity of Lactobacillus casei ΔRR12. RR12 controls the expression of the dlt operon, and its inactivation leads to a diminished teichoic acid D‐alanylation activity. To this end, a comparison of its response of ΔRR12 to low pH with the parental strain Lact. casei BL23 was carried out. Methods and Results The ability to induce an acid tolerance response (ATR), fatty acid (FA) composition and proteome changes induced in both strains in response to acid were investigated. Results obtained showed that both strains induce a growth‐phase‐dependent ATR. However, significant differences in the content of FAs and membrane‐associated proteins were detected. Conclusions The greater abundance of cytoplasmic proteins in the membrane fraction of the mutant strain ΔRR12 suggests an increased permeability of the cell membrane in this strain. Significance and Impact of the Study The analysis of the response to low pH of strain ΔRR12 indicated that the inactivation of TCS12 affected the content of FAs and proteins associated to the cell envelope. Increased abundance of cytoplasmic proteins suggested that low alanylation of teichoic acids affected the permeability of the cell membrane and possibly accounts for the acid sensitivity of strain ΔRR12.

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Section: Discussionmentioning

confidence: 65%

Characterization of the response to low pH ofLactobacillus caseiΔRR12, a mutant strain with low D-alanylation activity and sensitivity to low pH

et al. 2014

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“…The role of the GAPDH protein as an adhesin was confirmed in various bacterial species, including numerous bacteria from the genus Lactobacillus (Kainulainen et al, 2012;Kinoshita et al, 2012;Terrasse et al, 2015). The role of the TUFM protein in promoting the adhesion of bacteria (e.g.…”

Section: Detection Of Gapdh and Tufm In Extracellular Protein Fractiomentioning

confidence: 93%

Lactobacillus spp. belonging to the Casei group display a variety of adhesins

Konieczna

Olejnik-Schmidt

Schmidt

2018

Acta Sci Pol Technol Aliment

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Introduction. Adhesion of bacteria from the genus Lactobacillus to the gastrointestinal epithelium is, to a considerable degree, dependent on the interactions between adhesins found on the surface of bacterial cells and elements found within the epithelium. A significant role in these interactions is played by bacterial proteins exposed to the cell wall surface, which are capable of binding to molecules of substances comprising the extracellular matrix of the intestinal epithelium. Materials and methods.In order to analyze the extracellular proteome of intestinal bacteria in terms of the presence of cell adhesion molecules, a total of twenty strains from the Lactobacillus spp. group Casei were tested. The analyses were conducted using SDS PAGE, 2-D electrophoresis, Western blot and mass spectrometry. An experiment was also conducted to assess the adhesion capacity of the tested strains to cervical epithelial cells (HeLa).Results. The tested strains varied in their adhesion efficiency to HeLa cells, ranging from 0.5% to 29%. Using electrophoretic methods a total of 54 extracellular protein fractions were distinguished in these strains, additionally identifying potential adhesion molecules (e.g. a surface antigen of the NLP/P60 family and a small heat shock protein/chaperonin). Discussion. The identification of these proteins in the extracellular proteome of Latobacillus spp. isolates may suggest that they serve currently unknown functions on the cell surface, including those connected with the interactions between bacteria and the intestinal epithelium. Such analyses may provide insight into new factors promoting probiotic adhesion to various types of epithelial cells.

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“…To explore the impact of enzyme activity, GAPDH solution (2 mg/mL) was heated at 50 or 60°C for 1–3 h. GAPDH activity was evaluated according to the method described by Kinoshita et al . Briefly, 1 μL of glyceraldehyde‐3‐phosphate (47 mg/mL, Sigma‐Aldrich), 25 μL of NAD + (10 mM, Roche Diagnostics GmbH) and 175 μL of assay buffer (40 mM triethanolamine, 50 mM Na 2 HPO 4 , 0.2 mM ethylenediaminetetraacetic acid, 20 mM 2‐mercaptoethanol and 0.1% (v/v) Tween 20, pH 8.5) were added in 50 μL of each sample.…”

Section: Methodsmentioning

confidence: 99%

A novel moonlight function of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) for immunomodulation

et al. 2017

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an energy metabolism-related enzyme, which generates NADH in glycolysis. Our previous study revealed a novel role of exogenous GAPDH in the amelioration of lipopolysaccharide (LPS)-induced sepsis-related, severe acute lung injury (ALI) in mice. Here, we show the effect of extracellular GAPDH on the physiological functions of macrophages, which play an important role in the onset of sepsis and ALI. GAPDH has no effect on cell viability, while it strongly suppressed cell adhesion, spreading, and phagocytic function of LPS-stimulated macrophages. GAPDH treatment significantly reduced tumor necrosis factor (TNF)-α, while it induced interleukin (IL)-10 production from LPS-stimulated macrophages in a dose-dependent manner. It is noteworthy that heat inactivation of GAPDH lost its immunomodulatory activity. Correspondingly, NADH significantly inhibited TNF-α and enhanced IL-10 production with elevation of both M1/M2 macrophage markers. These data suggest that extracellular GAPDH induces intermediate M1/M2 macrophages for termination of inflammation, partly through its enzyme activity for generation of NADH. © 2017 BioFactors, 2017.

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Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH)

Cited by 15 publications

References 45 publications

Characterization of the response to low pH ofLactobacillus caseiΔRR12, a mutant strain with low D-alanylation activity and sensitivity to low pH

Characterization of the response to low pH ofLactobacillus caseiΔRR12, a mutant strain with low D-alanylation activity and sensitivity to low pH

Lactobacillus spp. belonging to the Casei group display a variety of adhesins

A novel moonlight function of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) for immunomodulation

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