Crystallization and Characterization of an Inflammatory Lectin Purified from the Seeds of Dioclea wilsonii
| Autor: | Raquel G. Benevides, Thaiz Batista Azevedo Rangel, Ana Maria Sampaio Assreuy, Kyria S. Nascimento, Rafael da Conceição Simões, Maria Júlia Barbosa Bezerra, Antonia Samia Fernandes do Nascimento, Alana de Freitas Pires, Plínio Delatorre, Alexandre Holanda Sampaio, Helton C. Silva, Amanda Uliana de Carvalho, Patricia Machado Bueno Fernandes, Celso Shiniti Nagano, Benildo Sousa Cavada, Bruno A.M. Rocha |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Erythrocytes
crystallization Neutrophils Molecular Sequence Data Ion chromatography Pharmaceutical Science Mannose Dioclea wilsonii Tandem mass spectrometry Article Analytical Chemistry lcsh:QD241-441 chemistry.chemical_compound Affinity chromatography lcsh:Organic chemistry Cell Movement Drug Discovery tandem mass spectrometry Metalloprotein Animals Humans Amino Acid Sequence Rats Wistar Physical and Theoretical Chemistry Conserved Sequence chemistry.chemical_classification Chromatography biology Protein Stability Organic Chemistry Lectin Rats Amino acid chemistry Biochemistry Chemistry (miscellaneous) Sephadex inflammation Seeds Dioclea biology.protein Molecular Medicine lectin Rabbits Inflammation Mediators Plant Lectins Sequence Alignment |
| Zdroj: | Molecules, Vol 16, Iss 6, Pp 5087-5103 (2011) Molecules Volume 16 Issue 6 Pages 5087-5103 |
| ISSN: | 1420-3049 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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