Mass Spectrometry and X-ray Diffraction Analysis of Two Crystal Types of Dioclea virgata Lectin: An Antinociceptive Protein Candidate to Structure/Function Analysis

• Published in: Applied biochemistry and biotechnology Vol. 164; no. 6; pp. 741 - 754
• Main Authors: Delatorre, Plínio, Rocha, Bruno A. M., Simões, Rafael C., Pereira-Júnior, Francisco N., Silva, Helton C., Bezerra, Eduardo Henrique S., Bezerra, Maria Julia B., Marinho, Emmanuel S., Gadelha, Carlos A. A., Santi-Gadelha, Tatiane, Farias, Daniel L., Assreuy, Ana Maria S., Marques-Domingos, Gabriela F. O., Nagano, Celso S., Cavada, Benildo S.
• Format: Journal Article
• Language: English
• Published: New York Humana Press Inc 01.07.2011; Springer; Springer Nature B.V
• Subjects: Amino Acid Sequence; Amino acids; Analgesics - chemistry; Analgesics - isolation & purification; Analgesics - pharmacology; Animals; Biochemistry; Biological and medical sciences; Biotechnology; Chemistry; Chemistry and Materials Science; Chromatography; Correlation coefficient; Crystallization; Crystals; Dioclea; Dioclea - chemistry; Edema - drug therapy; Fundamental and applied biological sciences. Psychology; Humans; Male; Mass Spectrometry; Mice; Molecular Sequence Data; Peptide Mapping; Plant Lectins - chemistry; Plant Lectins - isolation & purification; Plant Lectins - pharmacology; Proteins; Seeds - chemistry; Sequence Alignment; X-Ray Diffraction; Lectin; Tandem mass spectrometry; Antinociceptive; Crystallization; Mass spectrometry MS/MS; Crystals; X ray diffraction; Mass spectrometry; Protein; Structural analysis
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-011-9170-x

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.