Inhibition by Lactoferrin and<i>κ</i>-Casein Glycomacropeptide of Binding of<i>Cholera</i>Toxin to its Receptor

Kawasaki, Yuki; Isoda, Hiroko; Tanimoto, Morimasa; Dosako, Shun’ichi; Idota, Tadashi; Ahiko, Kenkichi

doi:10.1271/bbb.56.195

Cited by 110 publications

(61 citation statements)

References 5 publications

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“…CMP and CMP-derived peptides have been reported to have a variety of biological activities, such as suppression of gastric secretions (28), depression of platelet aggregation (2), inhibition of influenza virus hemagglutination (8), inhibition of cholera toxin binding (9), and immunomodulating activities (14). CMP has also been shown to incorporate into salivary pellicle and inhibit the adherence of Streptococcus mutans, the oral pathogen implicated in the development of dental caries (12,17,21,25,26).…”

mentioning

confidence: 99%

Kappacin, a Novel Antibacterial Peptide from Bovine Milk

Malkoski

Dashper

O'Brien-Simpson

et al. 2001

Antimicrob Agents Chemother

187

141

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Caseinomacropeptide (CMP) is a heterogeneous C-terminal fragment (residues 106 to 169) of bovine milk -casein composed of glycosylated and phosphorylated forms of different genetic variants. We have demonstrated that CMP has growth-inhibitory activity against the oral opportunistic pathogens Streptococcus mutans and Porphyromonas gingivalis and against Escherichia coli. CMP was fractionated using reversed-phase highperformance liquid chromatography (RP-HPLC), and each fraction was tested for activity against S. mutans in a 96-well-plate broth assay. Fractions were characterized by N-terminal sequence analysis and mass spectrometry. The active form of CMP was shown to be the nonglycosylated, phosphorylated -casein (residues 106 to 169) [-casein(106-169)], which we have designated kappacin. Endoproteinase Glu-C was used to hydrolyze CMP, and the generated peptides were separated using RP-HPLC and gel filtration-HPLC and then tested for activity against S. mutans. The peptide Ser(P) 149 -casein-A(138-158) was the only peptide generated by endoproteinase Glu-C digestion that exhibited growth-inhibitory activity. Peptides corresponding to the sequences of the inhibitory peptide Ser(P) 149 -casein-A(138-158) and its nonphosphorylated counterpartcasein-A(138-158) were chemically synthesized and tested for antibacterial activity. The synthetic Ser(P) 149-casein-A(138-158) displayed growth-inhibitory activity against S. mutans (MIC, 59 g/ml [26 M]). The nonphosphorylated peptide, however, did not inhibit growth at the concentrations tested, indicating that phosphorylation is essential for activity.The caseins are the most abundant bovine milk proteins, and there are four major types: ␣ s1 -, ␣ s2 -, ␤-, and -casein (23). All four caseins are phosphorylated on specific seryl residues, and in addition, -casein is glycosylated (4). -Casein is hydrolyzed by the enzyme chymosin between Phe 105 and Met 106 , generating two polypeptides: a hydrophobic N-terminal para--casein polypeptide -casein (residues 1 to 105) [-casein(1-105)] and a hydrophilic phosphorylated and glycosylated C-terminal polypeptide -casein(106-169), known as the caseinomacropeptide (CMP). CMP is heterogeneous and contains all the posttranslational modification sites (glycosylation and phosphorylation) of -casein. Six potential glycosylation sites have been identified on CMP (18), and up to five different carbohydrate moieties may be attached at each site (20). Three genetic variants of CMP have also been identified, originating from the precursors -casein A, B, and E, with variants A and B being the most common in bovine milk (15).CMP and CMP-derived peptides have been reported to have a variety of biological activities, such as suppression of gastric secretions (28), depression of platelet aggregation (2), inhibition of influenza virus hemagglutination (8), inhibition of cholera toxin binding (9), and immunomodulating activities (14). CMP has also been shown to incorporate into salivary pellicle and inhibit the adherence of Streptococcus mutans, the oral...

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mentioning

confidence: 99%

Kappacin, a Novel Antibacterial Peptide from Bovine Milk

Malkoski

Dashper

O'Brien-Simpson

et al. 2001

Antimicrob Agents Chemother

187

141

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“…I) Both toxins elevate intracellular cyclic AMP by activating adenylate cyclase in the target cells and then cause secretory diarrhea. L T and cholera toxin both use G M 1 ganglioside as a cell-surface receptor,2) and certain intestinal glycoproteins containing a terminal galactose residue (galactoproteins) also seem to be receptors for L T. 3 -6) The binding of L T to the receptors on intestinal epithelial cell surfaces must occur if it is to act as a toxin, so compounds with a sugar sequence similar or identical to GM 1 or the galactoprotein receptor may inhibit diarrhea by competition with the receptors for L T. Kawasaki et al 7) showed that lactoferrin and K-casein, which contain sugar chains similar to GM I, inhibit morphological changes caused by cholera toxin in CHO-K 1 cells. Otmess et al 8 ) found that GMI in milk inhibits fluid secretion caused by L T or cholera toxin in rabbit intestine, and Stoll et af.…”

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confidence: 99%

“…The ability of Lac-LA to inhibit the toxic activity of L Twas tested with CHO-K I cells (Dainippon Pharmaceutical Co., Ltd., Osaka, Japan) as described by Kawasaki et al 7) Lac-LA was serially diluted in Ham's FI2 medium supplemented with 1 % heat-inactivated fetal calf serum, mixed with an equal volume of L T dissolved at 6 ng/ml in the medium, and incubated for 30 min at 4C. The mixture (450 J.t!)…”

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confidence: 99%

Inhibition ofEscherichia coliHeat-labile Enterotoxin by an Amino Carbonyl Product, Lactose-α-lactalbumin

Shida

Takamizawa

Osawa

1996

Bioscience, Biotechnology, and Biochemistry

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The inhibition by lactose-~-Iactalbumin amino carbonyl product of Escherichia coli heat-labile enterotoxin was studied by GMt-ELISA and by assay with CHO-Kl cells. The product dosedependently inhibited the binding of the enterotoxin to GMI ganglioside and decreased the morphological change of CHO-KI cells caused by this toxin. The results suggest that this product may be a receptor analogue in the intestine.Key words: Escherichia coli heat-labile enterotoxin (L T); lactose-protein amino carbonyl product; receptor analogue Escherichia coli heat-labile enterotoxin (L T) type I is structurally and functionally similar to cholera toxin. I) Both toxins elevate intracellular cyclic AMP by activating adenylate cyclase in the target cells and then cause secretory diarrhea. L T and cholera toxin both use G M 1 ganglioside as a cell-surface receptor,2) and certain intestinal glycoproteins containing a terminal galactose residue (galactoproteins) also seem to be receptors for L T. 3 -6) The binding of L T to the receptors on intestinal epithelial cell surfaces must occur if it is to act as a toxin, so compounds with a sugar sequence similar or identical to GM 1 or the galactoprotein receptor may inhibit diarrhea by competition with the receptors for L T. Kawasaki et al. 7) showed that lactoferrin and K-casein, which contain sugar chains similar to GM I, inhibit morphological changes caused by cholera toxin in CHO-K 1 cells. Otmess et al. )found that GMI in milk inhibits fluid secretion caused by L T or cholera toxin in rabbit intestine, and Stoll et af. 9) reported that oral administration ofGM 1 adsorbed by charcoal reduced purging in patients with severe cholera.L T binds to proteins glycosylated with lactose by an amino carbonyl reaction through recognition of lactuloselysine (GalfJl4Fru-Lys), which is generated by Amadori rearrangement. 10) Lactose-protein amino carbonyl products may be receptor analogue that can inhibit diarrhea caused by L T. To evaluate this idea, we prepared lactose-~-Iactalbumin amino carbonyl products (Lac-LAs) with different degrees of glycosylation, and studied their ability to inhibit the binding of L T to GM I receptor and the morphological changes caused by LT in CHO-K I cells.L T (type I) was purified from a human strain of enterotoxigenic E. coli 240-3 by affinity chromatography on an immobilized D-galactose column (Pierce Chemical Co., Rockford, III., U.S.A.) by the method of Uesaka et al. II) Lac-LA was prepared as described by Matsuda et al. 12) Briefly, :x-lactalbumin and an equal weight of lactose were dissolved in distilled water and freeze-dried. The powder was kept at 65% relative humidity at 50 C for 2 or 5 days. The reaction product was dialyzed against distilled water to remove free sugar and then freeze-dried. The extent of glycosylation of Lac-LA samples was assayed by the furosine method 13) as described previously. I 0) lactuloselysine, an Amadori rearrangement compound anS1l1g from an amino carbonyl reaction between lactose and lysine residues.A microtiter-well binding ...

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“…It has been shown that the receptor is a oligosaccharide ganglioside GM1 [Van Heyningen, 1974], which is not identical to GMP, but other glycoproteins such as fetuin and glycophorin which have oligosaccharides similar to GMP inhibit cholera toxin [Sugii & Tsuji, 1990;Schengrund & Ringler, 1989]. Kawasaki et al [1992] have shown that GMP is capable of binding cholera toxin. Normal Chinese hamster ovary (CHO)-K1 cells are spherical.…”

Section: Biological Activities Ability To Bind Cholera Toxin and E Cmentioning

confidence: 99%

Healthy Multifunctional Spectra of Milk Glycoproteins and Their Fragments – a Review Article

Fayed¹

2012

Pol. J. Food Nutr. Sci.

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Inhibition by Lactoferrin andκ-Casein Glycomacropeptide of Binding ofCholeraToxin to its Receptor

Cited by 110 publications

References 5 publications

Kappacin, a Novel Antibacterial Peptide from Bovine Milk

Kappacin, a Novel Antibacterial Peptide from Bovine Milk

Inhibition ofEscherichia coliHeat-labile Enterotoxin by an Amino Carbonyl Product, Lactose-α-lactalbumin

Healthy Multifunctional Spectra of Milk Glycoproteins and Their Fragments – a Review Article

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