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

被引：19

作者：

Azevedo Rangel, Thaiz Batista ^{[1
]}

Sampaio Assreuy, Ana Maria ^{[2
]}

Pires, Alana de Freitas ^{[2
]}

de Carvalho, Amanda Uliana ^{[1
]}

Benevides, Raquel Guimaraes ^{[3
]}

Simoes, Rafael da Conceicao ^{[3
]}

da Silva, Helton Colares ^{[3
]}

Barbosa Bezerra, Maria Julia ^{[3
]}

Fernandes do Nascimento, Antonia Samia ^{[3
]}

do Nascimento, Kyria Santiago ^{[3
]}

Nagano, Celso Shiniti ^{[3
]}

Sampaio, Alexandre Holanda ^{[3
]}

Delatorre, Plinio ^{[4
]}

Matias da Rocha, Bruno Anderson ^{[3
]}

Bueno Fernandes, Patricia Machado ^{[1
]}

Cavada, Benildo Sousa ^{[3
]}

机构：

[1] Univ Fed Espirito Santo, Nucleo Biotecnol, Ctr Ciencias Saude, BR-29040090 Vitoria, ES, Brazil

[2] Univ Estadual Ceara, Inst Super Ciencias Biomed, BR-60455970 Fortaleza, Ceara, Brazil

[3] Univ Estadual Ceara, Dept Bioquim & Biol Mol, Lab Mol Biol Ativas, BR-60455970 Fortaleza, Ceara, Brazil

[4] Univ Fed Paraiba, Dept Biol Mol, Ctr Ciencias Exatas & Nat, BR-58051970 Joao Pessoa, Paraiba, Brazil

来源：

MOLECULES | 2011年 / 16卷 / 06期

关键词：

crystallization; Dioclea wilsonii; inflammation; lectin; tandem mass spectrometry; STIMULATION; PROTEINS; SEQUENCE;

D O I：

10.3390/molecules16065087

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

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 alpha (25,634 +/- 2 Da), beta (12,873 +/- 2 Da) and gamma (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 angstrom 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 angstrom. 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.

引用

页码：5087 / 5103

页数：17

共 45 条

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