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Purification and partial characterization of a new mannose/glucose‐specific lectin from Dialium guineense Willd seeds that exhibits toxic effect
Author(s) -
Bari Alfa U.,
Silva Helton C.,
Silva Mayara T. L.,
Pereira Júnior Francisco N.,
Cajazeiras João B.,
Sampaio Alexandre H.,
Leal Rodrigo B.,
Teixeira Edson H.,
Rocha Bruno A. M.,
Nascimento Kyria S.,
Nagano Celso S.,
Cavada Benildo S.
Publication year - 2013
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.2279
Subject(s) - mannose , chemistry , lectin , chromatography , sepharose , affinity chromatography , molecular mass , incubation , biochemistry , glucosamine , polyacrylamide gel electrophoresis , electrospray ionization , gel electrophoresis , electrophoresis , mass spectrometry , enzyme
A new mannose/glucose‐specific lectin, named DigL, was purified from seeds of Dialium guineense by a single step using a Sepharose 4b‐Mannose affinity chromatography column. DigL strongly agglutinated rabbit erythrocytes and was inhibited by d ‐mannose, d ‐glucose, and derived sugars, especially α‐methyl‐ d ‐mannopyranoside and N ‐acetyl‐ d ‐glucosamine. DigL has been shown to be a stable protein, maintaining its hemagglutinating activity after incubation at a wide range of temperature and pH values and after incubation with EDTA. DigL is a glycoprotein composite by approximately 2.9% of carbohydrates by weight. By sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis, the purified DigL exhibited an electrophoretic profile consisting of a broad band of 28–30 kDa. Analysis using electrospray ionization mass spectrometry indicated that purified DigL possesses a molecular average mass of 28 452 ± 2 Da and shows the presence of possible glycoforms. In addition, DigL exhibited an intermediary toxic effect on Artemia sp. nauplii, and this effect was both dependent on native structure and mediated by a carbohydrate‐binding site. Copyright © 2013 John Wiley & Sons, Ltd.