Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii
AbstractDwL, 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.
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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.Chicago/Turabian Style
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.