Binding of methylumbelliferyl N-acetyl-β-d-glucopyranoside and 4-methylumbelliferyl N, N′-diacetyl-β-chitobioside to rice lectin: studies by equilibrium dialysis and fluorescence quenching titrations

Poola, I. and Kella, N. K. D. (1986) Binding of methylumbelliferyl N-acetyl-β-d-glucopyranoside and 4-methylumbelliferyl N, N′-diacetyl-β-chitobioside to rice lectin: studies by equilibrium dialysis and fluorescence quenching titrations. BBA - General Subjects, 882 (1). pp. 12-17. ISSN 0304-4165

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Abstract

The binding of two fuorescent sugars 4-methylumbelliferyl N-actyl-β-d-glucopyranoside (MeUmbGlcNac) and 4-methylumbelliferyl N,N′-diacetyl-β-chitobioside (MeUmb(GlcNAc)2) to rice lectine was studied by equilibrium dialysis and extrinsic fluorescence titrations. Rice lectin contains four identical and independent carbohydrate binding sites per 36 000 Mr protein. Equilibrium dialysis experiments at 25°C gave an association constant value of (4.82 ± 0.32) · 104 M-1 whereas fluorescence quenching titrations at 15°C yielded a value of 8.17 +- 0.25) · 104 M-1 with MeUmbGlcNAc. The relative fluorescence quenching efficiency of the sugar increased with the saccharide chain lenght. The calculated quenching of fluorescence in both cases was 100%, indicating that the sugar binding site on the lectin is hydrophobic in nature. Binding constants and thermodynamic parameters of the binding of MeUmb glycosides were determined at 10, 15, 20 and 25°C. In both cases, binding constants decreased with increasing temperature, indicating that the binding of sugar to the lectin is exothermic in nature. The ΔH° and Δ° values for MeUmbGlcNAc-rice lectin interaction were -8.51 kcal·mol-1 and -7.07 e.u., respectively, and for that of MeUmb(GlcNAc)2-rice lectin the corresponding values were -11.89 kcal·mol-1 and -16.72 e.u.

Item Type: Article
Subjects: C Chemical Science > Biochemistry
Divisions: Department of > Biochemistry
Depositing User: Dhruva Kumar
Date Deposited: 10 May 2021 05:08
Last Modified: 10 May 2021 05:08
URI: http://eprints.uni-mysore.ac.in/id/eprint/13612

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