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

Publication date: September 2017
Source:International Journal of Biological Macromolecules, Volume 102
Author(s): David Alencar Araripe, Vanir Reis Pinto-Junior, Antonio Hadson Bastos Neco, Mayara Queiroz Santiago, Vinicius Jose Silva Osterne, Alana Freitas Pires, Claudia Figueiredo Lossio, Maria Gleiciane Queiroz Martins, Jorge Luiz Almeida Correia, Raquel Guimaraes Benevides, Rodrigo Bainy Leal, Ana Maria Sampaio Assreuy, Kyria Santiago Nascimento, Benildo Sousa Cavada
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.



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