title: Crystallization and characterization of an inflammatory lectin purified from the seeds of Dioclea wilsonii.
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.
authors.affiliation: Núcleo de Biotecnologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES, Brazil. thaizrangel@gmail.com
authors.forename: Thaiz Batista Azevedo
authors.initials: TB
authors.lastname: Rangel
authors.forename: Ana Maria Sampaio
authors.initials: AM
authors.lastname: Assreuy
authors.forename: Alana de Freitas
authors.initials: Ade F
authors.lastname: Pires
authors.forename: Amanda Uliana de
authors.initials: AU
authors.lastname: Carvalho
authors.forename: Raquel Guimarães
authors.initials: RG
authors.lastname: Benevides
authors.forename: Rafael da Conceição
authors.initials: Rda C
authors.lastname: Simões
authors.forename: Helton Colares da
authors.initials: HC
authors.lastname: Silva
authors.forename: Maria Júlia Barbosa
authors.initials: MJ
authors.lastname: Bezerra
authors.forename: Antonia Samia Fernandes do
authors.initials: AS
authors.lastname: Nascimento
authors.forename: Kyria Santiago do
authors.initials: KS
authors.lastname: Nascimento
authors.forename: Celso Shiniti
authors.initials: CS
authors.lastname: Nagano
authors.forename: Alexandre Holanda
authors.initials: AH
authors.lastname: Sampaio
authors.forename: Plínio
authors.initials: P
authors.lastname: Delatorre
authors.forename: Bruno Anderson Matias da
authors.initials: BA
authors.lastname: Rocha
authors.forename: Patricia Machado Bueno
authors.initials: PM
authors.lastname: Fernandes
authors.forename: Benildo Sousa
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D017124:Conserved Sequence; D003460:Crystallization; D052598:Dioclea / Q000737:chemistry*; D004912:Erythrocytes / Q000187:drug effects; D006801:Humans; D018836:Inflammation Mediators / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D055550:Protein Stability; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012639:Seeds / Q000737:chemistry*; D016415:Sequence Alignment
chemical_list: D018836:Inflammation Mediators; D037121:Plant Lectins