Purification of wheat germ agglutinin by affinity chromatography on a Sepharose-bound N-acetylglucosamine derivative

Lotan, Reuben; Gussin, Arnold E.S.; Lis, Halina; Sharon, Nathan

doi:10.1016/0006-291x(73)90763-8

Cited by 75 publications

(13 citation statements)

References 10 publications

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“…1B) and with activated but underivatized resin (not shown); this interaction is inhibited by chitin hydrolyzate. About 75% of the WGA applied to the columns can be recovered, as previously found with other ligand matrices [2,10]. The fact that both Con A and WGA bind quantitatively and reversibly to resin coupled with their respective haptens shows that the derivatization procedure can yield good matrices for affinity chromatography of lectins.…”

Section: Resultssupporting

confidence: 62%

A rapid procedure for derivatizing agarose with a variety of carbohydrates: Its use for affinity chromatography of lectins

Bloch

Burger

1974

FEBS Letters

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Section: Resultssupporting

confidence: 62%

A rapid procedure for derivatizing agarose with a variety of carbohydrates: Its use for affinity chromatography of lectins

Bloch

Burger

1974

FEBS Letters

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“…Wheat germ agglutinin was purified from commercial wheat germ by affinity chromatography [12]. Linear peptidoglycan (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…For relative inhibitory activity the hemagglutination inhibitory activity of N-acetyl-D-glucosamine was taken as 1. Apparent K, was calculated from the relative inhibitory activity using the K, for chitotriose (Ka = 2.2 x 104 M-') which was experimentally determined by us [21] and by Privat et al [26]. n.d. = not determined ghosts and normalized to GlcNAc content.…”

Section: Bacterial Agglutinationmentioning

confidence: 99%

“…From the relative inhibitory activity, the apparent association constants of the various inhibitors were calculated. The calculation was based on the binding constant for chitotriose, K, = 2.2 x 10" M which was measured by us [21] and by the group of Monsigny [26], using spectrofluorinietric titrations. The high apparent binding constants for the cell wall polymers are comparable to those calculated for the binding of wheat germ agglutinin to human erythrocytes [25].…”

Section: Nature Ofthe Interaction Between Wheat Germ Agglutinin and Pmentioning

confidence: 99%

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Interaction of Wheat‐Germ Agglutinin with Bacterial Cells and Cell‐Wall Polymers

Lotan

Sharon

Mirelman

1975

European Journal of Biochemistry

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Wheat germ agglutinin was found to agglutinate cells of Escherichia coli PAT 84, Micrococcus luteus, Staphylococcus aureus H, and of S. aureus 52A5, but not cells of S. aureus 52A2.Interaction of wheat germ agglutinin with a soluble linear peptidoglycan secreted by Micrococcus luteus and with the teichoic acid of S. aureus H was demonstrated by agar gel diffusion, quantitative precipitation and inhibition of hemagglutination of trypsinized rabbit erythrocytes. No interaction could be demonstrated with the teichoic acid from a phage-resistant mutant ( S . aureus 52A2) which lacks N-acetyl-D-glucosamine residues.All interactions were specifically inhibited by low concentrations of chitotriose (GlcNAcPl -+ 4GlcNAcPI + 4GlcNAc) and the bacterial cell wall tetrasaccharide, GlcNAcfil -+ 4MurNAcfil + 4GlcNAcjll + 4MurNAc. Hemagglutination-inhibition experiments showed that the linear peptidoglycan and the teichoic acid of S. aureus H were several thousand times more potent inhibitors of wheat germ agglutinin than was N-acetyl-D-glucosamine.Comparison of the efficiency of different saccharides in inhibition of hemagglutination and precipitation of polymers by wheat germ agglutinin, strongly suggests that secondary, non-specific interactions contribute to the binding of the lectin to the polymers.Many lectins agglutinate bacteria and precipitate bacterial polysaccharides specifically [ 1,2]. As early as 1936, Sumner and Howell [3] observed that concanavalin A agglutinated various Mycobacteria. This agglutination is at least in part due to the interaction of concanavalin A with an arabinogalactan present in mycobacterial cell walls [4]. Concanavalin A also precipitated the polyglycosylglycerol phosphate teichoic acid from Bacillus subtilis 168 [5].The lectin from snail (Helix pomatia) precipitated lipopolysaccharides of rough mutants of Salmonella typhimurium [6], Streptococcal group C polysaccharide, and the teichoic acid of Staphylococcus aureus 3528 in which nonreducing a-N-acetyl-D-glucosaminyl groups are present, but did not precipitate the teichoic acids of S. aureus A1 in which j-N-acetyl-D-glucosaminyl groups are attached instead [7,8].Interesting reports on the binding of phytohemagglutinin the nitrogen-fixing Rhizobia have recently appeared, and it has been suggested that these interactions may be responsible for the beneficial symbiotic relationship between the bacteria and the plants.Because of the growing interest in lectins, and the fact that they are useful for structural studies of complex saccharides [l, 21, we have investigated the interaction of wheat germ agglutinin, a lectin specific for N-acetyl-D-glucosamine residues [ll], with several bacteria and bacterial polymers. MATERIALS AND METHODS MaterialsN-Acetyl-D-glucosamine was a gift from PfiZer Inc. It was recrystallized from ethanol/acetone to give a melting point of 202 -203 "C. Chitotriose, (GlcNAc~1+4GlcNAc~1-+4GlcNAc) isolated from chitin, GlcNAcfil+4MurNAc and GlcNAcPl+4MurNAc-Pl+4GlcNAcP1-+4MurNAc from cell walls of MicroEur. J. Biochem. 55 (1...

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“…WGL was prepared according to Burger and Goldberg [2]; part of the preparation used in this experiment was purified by affinity chromatography [17] and was kindly donated by Dr. Nathan Sharon, Weizmann Institue of Science, Israel. Both preparations showed an identical precipitin reaction with a specific glycolipid and glycoprotein.…”

Section: Methodsmentioning

confidence: 99%

A Glycosphingolipid sharing reactivity with both wheat germ lectin and ‘carcinoembryonic antisera (gold)’: Partial identity of these reactive sites

1973

FEBS Letters

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Purification of wheat germ agglutinin by affinity chromatography on a Sepharose-bound N-acetylglucosamine derivative

Cited by 75 publications

References 10 publications

A rapid procedure for derivatizing agarose with a variety of carbohydrates: Its use for affinity chromatography of lectins

A rapid procedure for derivatizing agarose with a variety of carbohydrates: Its use for affinity chromatography of lectins

Interaction of Wheat‐Germ Agglutinin with Bacterial Cells and Cell‐Wall Polymers

A Glycosphingolipid sharing reactivity with both wheat germ lectin and ‘carcinoembryonic antisera (gold)’: Partial identity of these reactive sites

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