Purification, crystallization and preliminary X-ray analysis of a deletion mutant of a major buckwheat allergen

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Dec 1;65(Pt 12):1267-70. doi: 10.1107/S1744309109043127. Epub 2009 Nov 27.

Abstract

A 16 kDa buckwheat protein (BWp16) is a major allergen responsible for immediate hypersensitivity reactions including anaphylaxis. A deletion mutant of BWp16 (rBWp16DeltaN) was overproduced and purified and was shown to be immunologically active. A three-wavelength MAD data set was collected from a crystal of selenomethionine-labelled rBWp16DeltaN. The crystal belonged to the triclinic space group P1, with unit-cell parameters a = 28.39, b = 31.54, c = 32.20 A, alpha = 111.92, beta = 108.91, gamma = 98.74 degrees . One monomer was expected to be present in the asymmetric unit based on the calculated Matthews coefficient of 1.76 A(3) Da(-1).

MeSH terms

  • Allergens / chemistry*
  • Allergens / genetics
  • Base Sequence
  • Crystallography, X-Ray
  • DNA Primers / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Fagopyrum / chemistry
  • Fagopyrum / genetics
  • Fagopyrum / immunology*
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Plant Proteins / immunology*
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sequence Deletion

Substances

  • Allergens
  • DNA Primers
  • Plant Proteins
  • Recombinant Proteins