title: Mass spectrometry and X-ray diffraction analysis of two crystal types of Dioclea virgata lectin: an antinociceptive protein candidate to structure/function analysis.
abstract: The lectin from seeds of Dioclea virgata (DvirL) was purified in a single step affinity chromatography, sequenced by tandem mass spectrometry and submitted to crystallization and biological experiments. DvirL has a molecular mass of 25,412 ± 2 Da and the chains β and γ has 12,817 Da ± 2 and 12,612 Da ± 2, respectively. Primary sequence determination was assigned by tandem mass spectrometry and revealed a protein with 237 amino acids and 87% of identify with ConA. The protein crystals were obtained native and complexed with X-Man using vapor-diffusion method at a constant temperature of 293 K. A complete X-ray dataset was collected at 1.8 Å resolution. DvirL crystals were found to be orthorhombic, belonging to the space group I222, with a unit cell parameters a = 647.5 Å, b = 86.6 Å, c = 90.2 Å. Molecular replacement search found a solution with a correlation coefficient of 77.1% and an R(factor) of 44.6%. The present study also demonstrated that D. virgata lectin presents edematogenic and antinociceptive activities in rodents electing this protein as a candidate to structure/function analysis.
authors.affiliation: Departamento de Biologia Molecular, Universidade Federal da Paraíba, João Pessoa, Brazil.
authors.forename: Plínio
authors.initials: P
authors.lastname: Delatorre
authors.forename: Bruno A M
authors.initials: BA
authors.lastname: Rocha
authors.forename: Rafael C
authors.initials: RC
authors.lastname: Simões
authors.forename: Francisco N
authors.initials: FN
authors.lastname: Pereira-Júnior
authors.forename: Helton C
authors.initials: HC
authors.lastname: Silva
authors.forename: Eduardo Henrique S
authors.initials: EH
authors.lastname: Bezerra
authors.forename: Maria Julia B
authors.initials: MJ
authors.lastname: Bezerra
authors.forename: Emmanuel S
authors.initials: ES
authors.lastname: Marinho
authors.forename: Carlos A A
authors.initials: CA
authors.lastname: Gadelha
authors.forename: Tatiane
authors.initials: T
authors.lastname: Santi-Gadelha
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Farias
authors.forename: Ana Maria S
authors.initials: AM
authors.lastname: Assreuy
authors.forename: Gabriela F O
authors.initials: GF
authors.lastname: Marques-Domingos
authors.forename: Celso S
authors.initials: CS
authors.lastname: Nagano
authors.forename: Benildo S
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D000595:Amino Acid Sequence; D000700:Analgesics / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D003460:Crystallization; D052598:Dioclea / Q000737:chemistry*; D004487:Edema / Q000188:drug therapy; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D012639:Seeds / Q000737:chemistry; D016415:Sequence Alignment; D014961:X-Ray Diffraction
chemical_list: D000700:Analgesics; D037121:Plant Lectins