title: Purification and partial characterization of a new mannose/glucose-specific lectin from Dialium guineense Willd seeds that exhibits toxic effect.
abstract: 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.
authors.affiliation: BioMol-Group, Department of Biochemistry and Molecular Biology, Federal University of Ceara, PO Box 6043, 60440-970 Fortaleza, CE, Brazil.
authors.forename: Alfa U
authors.initials: AU
authors.lastname: Bari
authors.forename: Helton C
authors.initials: HC
authors.lastname: Silva
authors.forename: Mayara T L
authors.initials: MT
authors.lastname: Silva
authors.forename: Francisco N
authors.initials: FN
authors.lastname: Pereira Júnior
authors.forename: João B
authors.initials: JB
authors.lastname: Cajazeiras
authors.forename: Alexandre H
authors.initials: AH
authors.lastname: Sampaio
authors.forename: Rodrigo B
authors.initials: RB
authors.lastname: Leal
authors.forename: Edson H
authors.initials: EH
authors.lastname: Teixeira
authors.forename: Bruno A M
authors.initials: BA
authors.lastname: Rocha
authors.forename: Kyria S
authors.initials: KS
authors.lastname: Nascimento
authors.forename: Celso S
authors.initials: CS
authors.lastname: Nagano
authors.forename: Benildo S
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D000818:Animals; D001156:Artemia / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects; D007887:Fabaceae / Q000737:chemistry*; D005947:Glucose / Q000378:metabolism*; D006384:Hemagglutination / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D037241:Mannose-Binding Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity*; D013058:Mass Spectrometry; D008970:Molecular Weight; D009844:Oligosaccharides / Q000494:pharmacology; D011817:Rabbits; D012639:Seeds / Q000737:chemistry*; D013696:Temperature; D018675:Toxicity Tests
chemical_list: D037241:Mannose-Binding Lectins; D009844:Oligosaccharides; D005947:Glucose