Binding of simple carbohydrates and some N-acetyllactosamine-containing oligosaccharides to Erythrina cristagalli agglutinin as followed with a fluorescent indicator ligand

Boeck, Hilde De; Loontiens, Frank G.; Lis, Halina; Sharon, Nathan

doi:10.1016/0003-9861(84)90352-7

Cited by 75 publications

(33 citation statements)

References 22 publications

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“…With about -50 ( 10 kJ mol -1 , this is in good agreement with the calorimetrically determined enthalpy of -58 kJ mol -1 at 20°C (Table 1 and linear fit in Figure 2C). The observed association rates are in a range observed for many other protein-saccharide systems, even when much smaller oligo-and monosaccharides are involved (51)(52)(53). For an association reaction requiring rotational alignment and specific interactions of two macromolecules, the rate estimated theoretically lies between 0.5 × 10 6 and 5 × 10 6 s -1 M -1 (54).…”

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confidence: 60%

Enthalpic Barriers to the Hydrophobic Binding of Oligosaccharides to Phage P22 Tailspike Protein

et al. 2001

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The structural thermodynamics of the recognition of complex carbohydrates by proteins are not well understood. The recognition of O-antigen polysaccharide by phage P22 tailspike protein is a highly suitable model for advancing knowledge in this field. The binding to octa-and dodecasaccharides derived from Salmonella enteritidis O-antigen was studied by isothermal titration calorimetry and stoppedflow spectrofluorimetry. At room temperature, the binding reaction is enthalpically driven with an unfavorable change in entropy. A large change of -1.8 ( 0.2 kJ mol -1 K -1 in heat capacity suggests that the hydrophobic effect and water reorganization contribute substantially to complex formation. As expected from the large heat-capacity change, we found enthalpy-entropy compensation. The calorimetrically measured binding enthalpies were identical within error to van't Hoff enthalpies determined from fluorescence titrations. Binding kinetics were determined at temperatures ranging from 10 to 30°C. The second-order association rate constant varied from 1 × 10 5 M -1 s -1 for dodecasaccharide at 10°C to 7 × 10 5 M -1 s -1 for octasaccharide at 30°C. The first-order dissociation rate constants ranged from 0.2 to 3.8 s -1 . The Arrhenius activation energies were close to 50 and 100 kJ mol -1 for the association and dissociation reactions, respectively, indicating mainly enthalpic barriers. Despite the fact that this system is quite complex due to the flexibility of the saccharide, both the thermodynamic and kinetic data are compatible with a simple one-step binding model. Carbohydrate-protein interactions play an important role in many biological recognition processes. Compared to protein-protein interactions (1-4), the relation between structure and thermodynamics of carbohydrate binding is much less well understood, although there are some very well studied systems (5-14) and although the concept of empirical parameters for calculating thermodynamic data from the change in accessible surface area has been applied here, too (14-16). Generally, most carbohydrate-protein interactions are enthalpically driven and opposed by an unfavorable change in entropy (17,18). Most of the determined changes in heat capacity for these binding reactions with small oligosaccharides were small (<0.5 kJ mol -1 K -1 ) and negative. Enthalpy-entropy compensation for different ligands to the same protein and even for totally different systems was described (17). This was mainly interpreted to be due to solvent reorganization (19). This and other results led to the conclusion that release of solvent molecules from polyamphiphilic surfaces can drive binding reactions not only by an increase in entropy (hydrophobic effect) but also by a decrease in enthalpy (20-23). Structural and solution data for binding of monosaccharides and small oligosaccharides to lectins and transport proteins indicate a somewhat higher number of hydrogen bonds and a higher binding enthalpy per contact surface than in protein folding and protein-protein complexes ...

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confidence: 60%

Enthalpic Barriers to the Hydrophobic Binding of Oligosaccharides to Phage P22 Tailspike Protein

et al. 2001

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“…Among the Gal/GalNAc-speci®c lectins, PNA is the only one which does not bind GalNAc at all (Swamy et al, 1991;Young et al, 1984). WBAI, EcorL and SBA bind Gal and GalNAc, although the af®nities are not the same for the sugars in each case (De Boeck et al, 1984;Schwarz et al, 1991;Surolia et al, 1996;Young et al, 1984). GS4 does not bind monosaccharides, but Gal and GalNAc can occur at the primary site.…”

Section: Protein±carbohydrate Interactions and Sugar Speci®citymentioning

confidence: 99%

Structures of the complexes of peanut lectin with methyl-β-galactose and N-acetyllactosamine and a comparative study of carbohydrate binding in Gal/GalNAc-specific legume lectins

Ravishankar

Suguna

Surolia

et al. 1999

Acta Crystallogr D Biol Crystallogr

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The crystal structures of complexes of peanut lectin with methyl-β-galactose and N-acetyllactosamine have been determined at 2.8 and 2.7 Å, respectively. These, and the complexes involving lactose and the T-antigenic disaccharide reported previously, permit a detailed characterization of peanut-lectin–carbohydrate association and the role of water molecules therein. The water molecules in the combining site are substantially conserved in the four complexes. The role of interacting sugar hydroxyl groups, when absent, are often mimicked by ordered water molecules not only at the primary combining site, but also at the site of the second sugar ring. The similarity of peanut-lectin–sugar interactions with those in other galactose/N-acetylgalactosamine-specific lectins also extend to a substantial degree to water bridges. The comparative study provides a structural explanation for the exclusive specificity of peanut lectin for galactose at the monosaccharide level, compared with that of the other lectins for galactose as well as N-acetylgalactosamine. The complexes also provide a qualitative structural rationale for differences in the strengths of binding of peanut lectin to different sugars.

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“…A,,, readings were taken only for monitoring column effluents, using the optical factor of the very similar Erythrina cristagalli lectin, which is 1.53 mg ' cm2 (De Boeck et al, 1984).…”

Section: Protein Determinationmentioning

confidence: 99%

Mutational Studies of the Amino Acid Residues in the Combining Site of Erythrina corallodendron Lectin

Adar

Sharon

1996

European Journal of Biochemistry

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High-resolution X-ray crystallography of the complex of the Gal/GalNAc-specific Erythrinu corullodendron lectin with lactose identified the amino acid side chains that form contacts with the galactose moiety of the disaccharide. The contribution of these amino acids to the binding of different monosaccharides and oligosaccharides by the lectin was examined by site-directed mutagenesis.Replacement of Phel31, on which the galactose is stacked, by tyrosine, gave a mutant with the same hemagglutinating activity and carbohydrate specificity as the parent lectin, but replacement by alanine or valine resulted in loss of activity. Mutations of Ala88, Asp89, and Asn133 produced mutants that were also inactive whereas those of the other combining site residues, Tyr106, Ala218, and Gln219, were biologically active. None of the active mutants interacted with mannose or glucose. Thus, conti-ary to an earlier assumption, Ala218 is not responsible for the inability of E. corullodendron lectin to bind these sugars. Our findings also demonstrate that Gln219 is not involved in galactose binding in solution, even though this is implicated by the crystal data. Instead, our data suggest that Gln219 assists in the ligation of N-acetyllactosamine to the lectin, by interacting with the acetamide group of the disaccharide.Coinparison with other legume lectins specific for niannoselglucose, galactose, N-acetylgaiactosamine, L-fucose or N-acetylglucosamine, shows that only three of the combining site residues of E. corallodendron lectin occupy invariant positions both in their primary and tertiary structures. These residues are an aspartic acid and an asparagine corresponding to positions 89 and 133, respectively, in E. corallodendron lectin, and an aromatic residue, either phenylalanine (as Phel31 in this lectin), tyrosine or tryptophan. We therefore postulate that these three residues are essential for ligand binding by all such lectins, irrespective of their specificity.Keywords: legume lectins ; combining site ; mutagenesis ; contact residues.Legume lectins are members of a large family of structurally similar proteins with distinct carbohydrate specificities. Over 80 of these lectins have been isolated and characterized to varying extents. Almost all are comprised of two or four identical or nearly identical subunits, of 25-30 kDa each, held together by non-covalent forces (Jordan and Goldstein, 1994;Konami et al., 1995;Sharon and Lis, 1990). Each subunit contains tightly bound metal ions (one Ca" and one Mn") and possesses a single combining site to which different carbohydrates are bound with high specificity, although with low affinity, usually in the millimolar range. Many, but not all, are glycoproteins, with one or two N-linked glycans per subunit. In the primary structures of some 40 of the legume lectins that have been determined, 40% of the amino acid residues are invariant, or conservatively substituted. Moreover, the tertiary structures of nine of these lectins that have been solved by X-ray crystallography are superimposa...

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Binding of simple carbohydrates and some N-acetyllactosamine-containing oligosaccharides to Erythrina cristagalli agglutinin as followed with a fluorescent indicator ligand

Cited by 75 publications

References 22 publications

Enthalpic Barriers to the Hydrophobic Binding of Oligosaccharides to Phage P22 Tailspike Protein

Enthalpic Barriers to the Hydrophobic Binding of Oligosaccharides to Phage P22 Tailspike Protein

Structures of the complexes of peanut lectin with methyl-β-galactose and N-acetyllactosamine and a comparative study of carbohydrate binding in Gal/GalNAc-specific legume lectins

Mutational Studies of the Amino Acid Residues in the Combining Site of Erythrina corallodendron Lectin

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