Mass spectrometry and X-ray diffraction analysis of two crystal types of Dioclea virgata lectin: an antinociceptive protein candidate to structure/function analysis

Appl Biochem Biotechnol. 2011 Jul;164(6):741-54. doi: 10.1007/s12010-011-9170-x. Epub 2011 Feb 22.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Analgesics / chemistry*
  • Analgesics / isolation & purification
  • Analgesics / pharmacology
  • Animals
  • Crystallization
  • Dioclea / chemistry*
  • Edema / drug therapy
  • Humans
  • Male
  • Mass Spectrometry
  • Mice
  • Molecular Sequence Data
  • Peptide Mapping
  • Plant Lectins / chemistry*
  • Plant Lectins / isolation & purification
  • Plant Lectins / pharmacology
  • Seeds / chemistry
  • Sequence Alignment
  • X-Ray Diffraction

Substances

  • Analgesics
  • Plant Lectins