Binding of Disaccharides by Peanut Agglutinin as Studied by Ultraviolet Difference Spectroscopy

Neurohr, Klaus J.; Bundle, David R.; Young, N. Martin; Mantsch, Henry H.

doi:10.1111/j.1432-1033.1982.tb19768.x

Cited by 38 publications

(18 citation statements)

References 32 publications

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“…The related Gal-P-1,4-Glc linkage has also been studied in solution (18,19) with similar results. The reported solid state (20) conformation for p-lactose as determined by X-ray diffraction studies is + = 50" and $ = lo0, in qualitative agreement with the solution studies.…”

supporting

confidence: 56%

Syntheses of model oligosaccharides of biological significance. 9. Syntheses of trideuteriomethyl di-3,6-O-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-β-D-galactopyranoside: the I antigen branch-point trisaccharide and related disaccharides

Whitfield¹,

Ruzicka²,

Carver³

et al. 1987

Can. J. Chem.

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. Can. J. Chem. 65,693 (1987). ' The title trisaccharide, 13c, was synthesized, as well as its two component disaccharides, 10c and l l c . Four disaccharides, 3c, 4c, 5c, and 7c, were also prepared to serve as model compounds for the investigation of the 3-dimensional structure of more complex oligosaccharides. The P-1,3 linkage was formed in 75% yield by coupling 3,4,6-tri-0-acetyl-2-deoxy-2-phthalimido-P-D-glucopyranosyl bromide (9) with trideuteriomethyl2-O-benzoyl-4,6-benzylidene-~-~-galactopyranoside (2a), using silver trifluoromethanesulphonate as a promoter in the presence of the base 2,6-di-tert-butyl-4-methylpyridine. Using a silver zeolite as a promoter under a variety of conditions, both a-1,3 and P-1,3 linkages were formed. The P-1,6 linkages were prepared in high yield using 9 and silver zeolite with either trideuteriomethyl 2,3-di-0-benzoyl-P-D-galactopyranoside Chem. 65, 693 (1987). On a synthCtisC le trisaccharide mentionnC dans le titre (13c) ainsi que les deux disaccharides qui le foment (10c et l l c ) . On a aussi prCparC les quatre disaccharides 3c, 4c, 5c et 7c comme composCs modkles pour 1'Ctude de la structure tridimensionnelle d'oligosaccharides plus complexes. On a form6 la liaison P-1,3 avec un rendement de 75% en procCdant au couplage du bromure de tri-0-acttyl-3,4,6 dCoxy-2 phtalimido-2 P-D-glycopyrannosyle (9) avec le 0-benzoyl-2 benzylidkne-4,6 P-D-galactopyrannoside de trideut6romCthyle (2a), sous l'influence du trifluoromCthanesulfonate d'argent comme promoteur de la reaction et en prksence de di-tert-butyl-2,6 mCthyl-4 pyridine comme base. Si on utilise le zeolite d'argent comme promoteur, sous diverses conditions, il y a formation des liaisons a-1,3 ainsi que p-1,3. On a prCparC les liaisons P-1 ,6 avec de bons rendements en faisant rCagir le composC 9, en prCsence de zColite d'argent comme promoteur, avec soit le di-0-benzoyl-2,3 P-D-galactopyrranoside de trideutCromCthyle (86) soit le 0-benzoyl-2 0-(tri-0-acCtyl-3,4,6 dCoxy-2 phtalimido-2 P-D-g1~~0pyrann0~yl)-3 P-D-galactopyrannoside de trideutCromCthyle ( l l b ) . On a utilisC le bromure de tri-0-benzyl-2,3,4 a-L-fucopyrannosyle (lb) pour la fucosylation, catalysCe par les ions halogknures, du composC 2a ou de son isomkre 0-benzoyle-3 (26). Les glycosylations des bromures de di-0-acCtyl-3,6 0-(tCtra-0-acCtyl-2,3,4,6 P-D-gala~topyranno~y1)-4 dCoxy-2 phtalimido-2 P-D-glucopyrannosyle (66) et de 0-(tetra-0-acCtyl-2,3,4,6 P-D-galactopyrannosy1)-4 tri-0-acCtyl-2,3,6 a -D -g l~~~p y r a n n~~y l e (50) par l'heptadeutCro-isopropanol se font sous l'influence du zeolite d'argent comme promoteur.[Traduit par la revue]As part of our program of studies of complex oligosaccharides, in particular those structures related to neoplasia (I), we have synthesized trisaccharide 13c and several related disaccharides 3c, 4c, 5c, 7c, and 10c as their P-glycosides. These anomerically pure oligosaccharides are suitable for the elucidation of their 3-dimensional structures by nrnr spectroscopic techniques (2).1i is our that to understand the biol...

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supporting

confidence: 56%

Syntheses of model oligosaccharides of biological significance. 9. Syntheses of trideuteriomethyl di-3,6-O-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-β-D-galactopyranoside: the I antigen branch-point trisaccharide and related disaccharides

Whitfield¹,

Ruzicka²,

Carver³

et al. 1987

Can. J. Chem.

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“…Purqication of the protein Peanut agglutinin was purified from buffer extracts of delipidated peanut meal by affinity chromatography on a lactose-substituted agarose matrix [3]. The initial PNA sample was material used for spectroscopic studies [3,4,7, 81; subsequent samples were prepared identically from the same batch of peanuts.…”

Section: Methodsmentioning

confidence: 99%

“…The initial PNA sample was material used for spectroscopic studies [3,4,7, 81; subsequent samples were prepared identically from the same batch of peanuts.…”

Section: Purqication Of the Proteinmentioning

confidence: 99%

“…The lectin recognises both a-and P-galactosides, and the disaccharide of highest known affinity is the T-antigen structure Gal(b1-3)GalNAc [2] (GalNAc, N-acetyl-D-galactosamine). Detailed studies of the interactions of PNA with monosaccharides and disaccharides have been made by UV difference [3,4], fluorescence [5,61 and NMR [7, 81 spectroscopy, and a model of the binding process has been proposed [8]. PNA is therefore one of the best characterized lectins in its functional aspects, but far less is known about its structure.…”

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confidence: 99%

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The amino acid sequence of peanut agglutinin

Young

Johnston

Watson

1991

European Journal of Biochemistry

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The amino acid sequence of peanut (Arachis hypogaea) agglutinin was determined from three major fragments obtained by mild acid cleavage at Asp-Pro peptide bonds. The sequence of 236 amino acids has residues identical to those that form the metal-binding site and the hydrophobic pocket in concanavalin A and other lectins, although the overall similarity is only 42%. In the segments of peanut agglutinin that correspond to the four loops that form the carbohydrate-binding site in concanavalin A and favin, several central residues are homologous, while others show changes to smaller side chains, such as Tyr --f Gly. The carbohydrate-binding site of peanut agglutinin may therefore have a similar peptide-backbone architecture, but form a considerably more open cleft.Peanut agglutinin (PNA) is a legume seed lectin that is specific for D-galactose residues at non-reducing terminal positions of glycoconjugates [l]. This specificity has made it useful for histochemical and cell-surface studies. The lectin recognises both a-and P-galactosides, and the disaccharide of highest known affinity is the T-antigen structure Gal(b1-3)GalNAc [2] (GalNAc, N-acetyl-D-galactosamine). Detailed studies of the interactions of PNA with monosaccharides and disaccharides have been made by UV difference [3,4], fluorescence [5, 61 and NMR [7, 81 spectroscopy, and a model of the binding process has been proposed [8]. PNA is therefore one of the best characterized lectins in its functional aspects, but far less is known about its structure. It is a tetramer of subunits of molecular mass 27 kDa [9], each with one bindingsite [3], and is not a glycoprotein [I]. PNA has been successfully crystallized for X-ray studies in two laboratories [lo, 1 I].A partial amino acid sequence was reported by Lauwereys et al. [I21 who encountered difficulties with multiple genetic forms and poorly soluble peptides. The former are to be expected in view of the multiple bands seen in isoelectric focussing and chromatofocussing experiments [13,14]. We have reinvestigated the amino acid sequence of PNA and report a complete sequence together with data on the genetic variation at some positions. The sequences of the peptide loops in the putative carbohydrate-binding-site region are discussed in relation to the X-ray results for the D-mannose-specific lectins concanavalin A Abbreviations. PNA, peanut agglutinin; GalNAc, N-acetyl-Dgalactosamine. MATERIALS AND METHODS Purqication of the proteinPeanut agglutinin was purified from buffer extracts of delipidated peanut meal by affinity chromatography on a lactose-substituted agarose matrix [3]. The initial PNA sample was material used for spectroscopic studies [3,4,7, 81; subsequent samples were prepared identically from the same batch of peanuts. Chemical cleavageThe protein was cleaved at Asp-Pro peptide bonds by limited acid hydrolysis [19]. PNA (22 mg) was dissolved in 2.28 ml 88% formic acid, 1.72 ml water was added and the solution was incubated at 37°C for 96 h. The digest was diluted and freeze-dried. The fragments...

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“…3,4 However, it cross-reacted with related ␤-galactosides such as ␣-lactose, Gal␤1-4Glc␣, and Gal␤1-3GlcNAc␤. 5 Normal colorectal tissues did not show any or only slight reactivity, 6 -12 whereas carcinomas 6 -11,13 were stained in the majority of the cases. In adenomas, PNA binding also could be observed.…”

mentioning

confidence: 99%

Thomsen‐Friedenreich antigen presents as a prognostic factor in colorectal carcinoma

Baldus

Zirbes

Hanisch

et al. 2000

Cancer

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Binding of Disaccharides by Peanut Agglutinin as Studied by Ultraviolet Difference Spectroscopy

Cited by 38 publications

References 32 publications

Syntheses of model oligosaccharides of biological significance. 9. Syntheses of trideuteriomethyl di-3,6-O-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-β-D-galactopyranoside: the I antigen branch-point trisaccharide and related disaccharides

Syntheses of model oligosaccharides of biological significance. 9. Syntheses of trideuteriomethyl di-3,6-O-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-β-D-galactopyranoside: the I antigen branch-point trisaccharide and related disaccharides

The amino acid sequence of peanut agglutinin

Thomsen‐Friedenreich antigen presents as a prognostic factor in colorectal carcinoma

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