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• Rangel, T
• Assreuy, A
• Pires, A
• Carvalho, A
• Benevides, R
• Simões, R
• Silva, H
• Bezerra, M
• Nascimento, A
• Nascimento, K
• Nagano, C
• Sampaio, A
• Delatorre, P
• Rocha, B
• Fernandes, P
• Cavada, B
• [+] on Google Scholar
• Rangel, T
• Assreuy, A
• Pires, A
• Carvalho, A
• Benevides, R
• Simões, R
• Silva, H
• Bezerra, M
• Nascimento, A
• Nascimento, K
• Nagano, C
• Sampaio, A
• Delatorre, P
• Rocha, B
• Fernandes, P
• Cavada, B
• [+] on PubMed
• Rangel, T
• Assreuy, A
• Pires, A
• Carvalho, A
• Benevides, R
• Simões, R
• Silva, H
• Bezerra, M
• Nascimento, A
• Nascimento, K
• Nagano, C
• Sampaio, A
• Delatorre, P
• Rocha, B
• Fernandes, P
• Cavada, B

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Molecules 2011, 16(6), 5087-5103; doi:10.3390/molecules16065087

Article

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

Thaiz Batista Azevedo Rangel ¹ , Ana Maria Sampaio Assreuy ² , Alana de Freitas Pires ² , Amanda Uliana de Carvalho ¹ , Raquel Guimarães Benevides ³ , Rafael da Conceição Simões ³ , Helton Colares da Silva ³ , Maria Júlia Barbosa Bezerra ³ , Antonia Samia Fernandes do Nascimento ³ , Kyria Santiago do Nascimento ³ , Celso Shiniti Nagano ³ , Alexandre Holanda Sampaio ³ , Plínio Delatorre ⁴ , Bruno Anderson Matias da Rocha ³ , Patricia Machado Bueno Fernandes ^1,*  and Benildo Sousa Cavada ^3,*

¹ Núcleo de Biotecnologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES 29040-090, Brazil ² Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará, P. O. Box 6043, 60455-970 Fortaleza, Ceará, Brazil ³ Laboratório de Moléculas Biologicamente Ativas, Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, P. O. Box 6043, 60455-970 Fortaleza, Ceará, Brazil ⁴ Departamento de Biologia Molecular, Centro de Ciências Exatas e da Natureza - Campus I, Universidade Federal da Paraíba,Caixa Postal 5009, 58051-970, João Pessoa, PB, Brazil

* Authors to whom correspondence should be addressed.

Received: 14 April 2011; in revised form: 7 June 2011 / Accepted: 17 June 2011 / Published: 20 June 2011

Download PDF Full-Text [699 KB, uploaded 20 June 2011 14:44 CEST]

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.

Keywords: crystallization; Dioclea wilsonii; inflammation; lectin; tandem mass spectrometry

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