Abstract
Using a polyclonal antibody raised against a highly conserved sequence of 38 amino acids containing the activation site (VTDSAAGAT) common to mammalian and yeast alkaline phosphatases (AP), we identified in decapsidated Saccharomyces boulardii a protein phosphatase detected by autoradiography as a single signal (63 kD). Using an affinity chromatography column, the protein phosphatase could be concentrated 39.1-fold and presented as a doublet of two subunits. Compared with rat and bovine purified intestinal AP, the enzyme from S. boulardii had a greater ability to dephosphorylate the lipopolysaccharide (LPS) of Escherichia coli 055B5. When tested in vivo, intraperitoneal injection of intact LPS to rats produced, after 9 h, 100 ng/mL of circulating tumor necrosis factor-α with inflammatory lesions and apoptotic bodies in the liver and the heart, whereas rats injected with partially dephosphorylated LPS produced only 40 ng/mL tumor necrosis factor-α without organic lesions. In conclusion, S. boulardii is able to inhibit toxicity of E. coli surface endotoxins by the release of a protein phosphatase exhibiting a great capacity of dephosphorylation.
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Abbreviations
- BBM:
-
brush border membrane
- IAP:
-
intestinal alkaline phosphatase
- LPS:
-
lipopolysaccharide
- PMSF:
-
phenylmethanesulfonylfluoride
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The authors thank Bernard Hublot and Paul Bernasconi for helpful technical comments and Christiane Neefs-Neirinck for typing the manuscript.
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Financial support and technical assistance for this study provided by Biocodex, Gentilly, France.
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Buts, JP., Dekeyser, N., Stilmant, C. et al. Saccharomyces boulardii Produces in Rat Small Intestine a Novel Protein Phosphatase that Inhibits Escherichia coli Endotoxin by Dephosphorylation. Pediatr Res 60, 24–29 (2006). https://doi.org/10.1203/01.pdr.0000220322.31940.29
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DOI: https://doi.org/10.1203/01.pdr.0000220322.31940.29
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