title: Partial characterization and immobilization in CNBr-activated Sepharose of a native lectin from Platypodium elegans seeds (PELa) and comparative study of edematogenic effect with the recombinant form.
abstract: The lectin from Platypodium elegans seeds (PELa) was purified by affinity chromatography in a mannose-agarose column. The lectin agglutinated rabbit erythrocytes and the agglutinating effect was inhibited by previous incubation with the glycoprotein fetuin, along with N-acetyl-d-glucosamine, D-mannose and its derivatives. The lectin maintained complete activity in temperatures ranging from 40 to 60°C and pH values ranging from 9 to 10. As a glycoprotein, PELa has a carbohydrate content of 2.2%, and its activity requires divalent cations such as Ca2+and Mn2+. Based on SDS-PAGE, PELa displays a profile similar to that of other Dalbergieae lectins with the main chain of molecular mass around 30kDa and two subunits of 19kDa and 10 kDa each. Two-dimensional (2D) electrophoresis revealed the presence of isoforms with different isoelectric points, and high-performance size exclusion chromatography (HPSEC) was performed to confirm the purity of the sample. The lectin was immobilized in CNBr-activated Sepharose 4B and successfully captured fetuin in solution, demonstrating that this lectin remains active and capable of binding carbohydrates. PELa showed effects different from those of its recombinant form in both pro- and anti-inflammatory tests.
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: David Alencar
authors.initials: DA
authors.lastname: Araripe
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Vanir Reis
authors.initials: VR
authors.lastname: Pinto-Junior
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Antonio Hadson Bastos
authors.initials: AHB
authors.lastname: Neco
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Mayara Queiroz
authors.initials: MQ
authors.lastname: Santiago
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Vinicius Jose Silva
authors.initials: VJS
authors.lastname: Osterne
authors.affiliation: Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brazil.
authors.forename: Alana Freitas
authors.initials: AF
authors.lastname: Pires
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Claudia Figueiredo
authors.initials: CF
authors.lastname: Lossio
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Maria Gleiciane Queiroz
authors.initials: MGQ
authors.lastname: Martins
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Jorge Luiz Almeida
authors.initials: JLA
authors.lastname: Correia
authors.affiliation: Universidade Estadual de Feira de Santana (UEFS), Feira de Santana, Bahia, Brazil.
authors.forename: Raquel Guimaraes
authors.initials: RG
authors.lastname: Benevides
authors.affiliation: Universidade Federal de Santa Catarina (UFSC), Florianópolis, Santa Catarina, Brazil.
authors.forename: Rodrigo Bainy
authors.initials: RB
authors.lastname: Leal
authors.affiliation: Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brazil.
authors.forename: Ana Maria Sampaio
authors.initials: AMS
authors.lastname: Assreuy
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Kyria Santiago
authors.initials: KS
authors.lastname: Nascimento
authors.affiliation: Universidade Federal do Ceará (UFC), Fortaleza, Ceará, Brazil. Electronic address: bscavada@ufc.br.
authors.forename: Benildo Sousa
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D000818:Animals; D004487:Edema / Q000139:chemically induced*; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology; D007887:Fabaceae / Q000737:chemistry*; D006384:Hemagglutination / Q000187:drug effects; D055767:Immobilized Proteins / Q000737:chemistry / Q000494:pharmacology*; D008297:Male; D037121:Plant Lectins / Q000737:chemistry / Q000494:pharmacology*; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology*; D012639:Seeds / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry*
chemical_list: D055767:Immobilized Proteins; D037121:Plant Lectins; D011994:Recombinant Proteins; D012685:Sepharose
keywords: Immobilization; Inflammation; PELa