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

Molecules

Thaiz Batista Azevedo Rangel¹  Ana Maria Sampaio Assreuy¹  Alana de Freitas Pires¹  Amanda Uliana de Carvalho¹  Raquel Guimarães Benevides¹  Rafael da Conceição Simões¹  Helton Colares da Silva¹  Maria Júlia Barbosa Bezerra¹  Antonia Samia Fernandes do Nascimento¹  Kyria Santiago do Nascimento¹  Celso Shiniti Nagano¹  Alexandre Holanda Sampaio¹  Plínio Delatorre¹  Bruno Anderson Matias da Rocha¹  Patricia Machado Bueno Fernandes¹ 

[1] ¹Núcleo de Biotecnologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES 29040-090, Brazil

关键词: crystallization;    Dioclea wilsonii;    inflammation;    lectin;    tandem mass spectrometry;   

DOI  :  10.3390/molecules16065087

来源: mdpi

【 摘 要 】

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.

【 授权许可】

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