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Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii
Núcleo de Biotecnologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES 29040-090, Brazil
Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará, P. O. Box 6043, 60455-970 Fortaleza, Ceará, Brazil
Laboratório de Moléculas Biologicamente Ativas, Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, P. O. Box 6043, 60455-970 Fortaleza, Ceará, Brazil
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
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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Rangel, T.B.A.; Assreuy, A.M.S.; Pires, A.F.; Carvalho, A.U.; Benevides, R.G.; Simões, R.C.; Silva, H.C.; Bezerra, M.J.B.; Nascimento, A.S.F.; Nascimento, K.S.; Nagano, C.S.; Sampaio, A.H.; Delatorre, P.; Rocha, B.A.M.; Fernandes, P.M.B.; Cavada, B.S. Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii. Molecules 2011, 16, 5087-5103.
Rangel TBA, Assreuy AMS, Pires AF, Carvalho AU, Benevides RG, Simões RC, Silva HC, Bezerra MJB, Nascimento ASF, Nascimento KS, Nagano CS, Sampaio AH, Delatorre P, Rocha BAM, Fernandes PMB, Cavada BS. Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii. Molecules. 2011; 16(6):5087-5103.
Rangel, Thaiz Batista Azevedo; Assreuy, Ana Maria Sampaio; Pires, Alana de Freitas; Carvalho, Amanda Uliana de; Benevides, Raquel Guimarães; Simões, Rafael da Conceição; Silva, Helton Colares da; Bezerra, Maria Júlia Barbosa; Nascimento, Antonia Samia Fernandes do; Nascimento, Kyria Santiago do; Nagano, Celso Shiniti; Sampaio, Alexandre Holanda; Delatorre, Plínio; Rocha, Bruno Anderson Matias da; Fernandes, Patricia Machado Bueno; Cavada, Benildo Sousa. 2011. "Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii." Molecules 16, no. 6: 5087-5103.