Molecules (Jun 2011)

Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii

  • Ana Maria Sampaio Assreuy,
  • Alana de Freitas Pires,
  • Amanda Uliana de Carvalho,
  • Raquel Guimarães Benevides,
  • Rafael da Conceição Simões,
  • Helton Colares da Silva,
  • Maria Júlia Barbosa Bezerra,
  • Antonia Samia Fernandes do Nascimento,
  • Kyria Santiago do Nascimento,
  • Celso Shiniti Nagano,
  • Alexandre Holanda Sampaio,
  • Plínio Delatorre,
  • Bruno Anderson Matias da Rocha,
  • Patricia Machado Bueno Fernandes,
  • Benildo Sousa Cavada,
  • Thaiz Batista Azevedo Rangel

DOI
https://doi.org/10.3390/molecules16065087
Journal volume & issue
Vol. 16, no. 6
pp. 5087 – 5103

Abstract

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DwL, a lectin extracted from the seeds of Dioclea wilsonii, is a metalloprotein with strong agglutinating activity against rabbit and ABO erythrocytes, inhibited by glucose and mannose. DwL was purified by affinity chromatography on a Sephadex G-50 column and ion exchange chromatography on a HiTrap SP XL column. SDS-PAGE revealed three electrophoretic bands corresponding to the α (25,634 ± 2 Da), β (12,873 ± 2 Da) and γ (12,779 ± 2 Da) chains. Protein sequencing was done by Tandem Mass Spectrometry. The primary sequence featured 237 amino acids and was highly homologous to other reported Diocleinae lectins. A complete X-ray dataset was collected at 2.0 Å for X-Man-complexed DWL crystals produced by the vapor diffusion method. The crystals were orthorhombic and belonged to the space group I222, with the unit-cell parameters a = 59.6, b = 67.9 and c = 109.0 Å. DWL differed in potency from other ConA-like lectins and was found to induce neutrophil migration in rats, making it particularly useful in structural/functional studies of this class of proteins.

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