1. Crystallization and characterization of an inflammatory lectin purified from the seeds of Dioclea wilsonii.
- Author
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Rangel TB, Assreuy AM, Pires Ade F, Carvalho AU, Benevides RG, Simões Rda C, Silva HC, Bezerra MJ, Nascimento AS, Nascimento KS, Nagano CS, Sampaio AH, Delatorre P, Rocha BA, Fernandes PM, and Cavada BS
- Subjects
- Amino Acid Sequence, Animals, Cell Movement drug effects, Conserved Sequence, Crystallization, Erythrocytes drug effects, Humans, Inflammation Mediators isolation & purification, Inflammation Mediators pharmacology, Molecular Sequence Data, Neutrophils drug effects, Plant Lectins isolation & purification, Plant Lectins pharmacology, Protein Stability, Rabbits, Rats, Rats, Wistar, Sequence Alignment, Dioclea chemistry, Inflammation Mediators chemistry, Plant Lectins chemistry, Seeds chemistry
- Abstract
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.
- Published
- 2011
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