title: Purification, partial characterization and immobilization of a mannose-specific lectin from seeds of Dioclea lasiophylla mart.
abstract: Lectin from the seeds of Dioclea lasiophylla (DlyL) was purified in a single step by affinity chromatography on a Sephadex® G-50 column. DlyL strongly agglutinated rabbit erythrocytes and was inhibited by monosaccharides (D-mannose and α-methyl-D-mannoside) and glycoproteins (ovalbumin and fetuin). Similar to other Diocleinae lectins, DlyL has three chains, α, β and γ, with mass of 25,569 ± 2, 12,998 ± 1 and 12,588 ± 1 Da, respectively, and has no disulfide bonds. The hemagglutinating activity of DlyL was optimal in pH 8.0, stable at a temperature of 70 °C and decreased in EDTA solution, indicating that lectin activity is dependent on divalent metals. DlyL exhibited low toxicity on Artemia sp. nauplii, but this effect was dependent on the concentration of lectin in solution. DlyL immobilized on cyanogen bromide-activated Sepharose® 4B bound 0.917 mg of ovalbumin per cycle, showing the ability to become a tool for glycoproteomics studies.
authors.affiliation: Laboratory of Biologically Active Molecules (Biomol-Lab), Department of Biochemistry and Molecular Biology, Federal University of Ceará, Av. Humberto Monte s/n, Bloco 907, Lab. 1075, Campus do Pici, Fortaleza-CE 60440-970, Brazil. juniorreis4@hotmail.com
authors.forename: Vanir Reis
authors.initials: VR
authors.lastname: Pinto-Júnior
authors.forename: Mayara Queiroz
authors.initials: MQ
authors.lastname: de Santiago
authors.forename: Vinícius José da Silva
authors.initials: VJ
authors.lastname: Osterne
authors.forename: Jorge Luis Almeida
authors.initials: JL
authors.lastname: Correia
authors.forename: Francisco Nascimento
authors.initials: FN
authors.lastname: Pereira-Júnior
authors.forename: João Batista
authors.initials: JB
authors.lastname: Cajazeiras
authors.forename: Mayron Alves
authors.initials: MA
authors.lastname: de Vasconcelos
authors.forename: Edson Holanda
authors.initials: EH
authors.lastname: Teixeira
authors.forename: Antônia Sâmia Fernandes
authors.initials: AS
authors.lastname: do Nascimento
authors.forename: Thaiz Batista Azevedo Rangel
authors.initials: TB
authors.lastname: Miguel
authors.forename: Emilio de Castro
authors.initials: Ede C
authors.lastname: Miguel
authors.forename: Alexandre Holanda
authors.initials: AH
authors.lastname: Sampaio
authors.forename: Kyria Santiago
authors.initials: KS
authors.lastname: do Nascimento
authors.forename: Celso Shiniti
authors.initials: CS
authors.lastname: Nagano
authors.forename: Benildo Sousa
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D000818:Animals; D001156:Artemia; D002614:Chelating Agents / Q000737:chemistry; D002846:Chromatography, Affinity; D052598:Dioclea / Q000737:chemistry*; D004492:Edetic Acid / Q000737:chemistry; D004912:Erythrocytes / Q000187:drug effects; D006384:Hemagglutination; D006388:Hemagglutinins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007928:Lethal Dose 50; D037241:Mannose-Binding Lectins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010047:Ovalbumin / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D011485:Protein Binding; D011817:Rabbits; D012639:Seeds / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry
chemical_list: D002614:Chelating Agents; D006388:Hemagglutinins; D037241:Mannose-Binding Lectins; D010936:Plant Extracts; D010047:Ovalbumin; D012685:Sepharose; D004492:Edetic Acid