Reexamination of the carbohydrate binding stoichiometry of lima bean lectin.

Abstract:

:The carbohydrate binding stoichiometry of lima bean lectin component III was reexamined using equilibrium dialysis and quantitative affinity chromatography following limited chemical modification. Equilibrium dialysis employing methyl[2-14C]benzamido-2-deoxy-alpha-D-galactopyranoside as ligand demonstrated that the lectin tetramer bound 4 mol of sugar with Kassoc = 1.44 +/- 0.13 X 10(3) M-1 (T = 5 degrees C, pH 7.0, ionic strength 0.1). The previous report of two sites/tetramer [Bessler, W. and Goldstein, I. J. (1974) Arch. Biochem. Biophys. 165, 444] appears to be the result of partial inactivation of the lectin due to oxidation of essential thiol groups. Following limited chemical modification of the thiol groups by methyl methanethiosulfonate, multiple intermediate forms with reduced affinity for Synsorb A were obtained. The number and hemagglutinating activities of these intermediates provided further support for the presence of four carbohydrate binding sites on lima bean lectin component III.

journal_name

Arch Biochem Biophys

authors

Roberts DD,Goldstein IJ

doi

10.1016/0003-9861(84)90113-9

subject

Has Abstract

pub_date

1984-04-01 00:00:00

pages

316-20

issue

1

eissn

0003-9861

issn

1096-0384

pii

0003-9861(84)90113-9

journal_volume

230

pub_type

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