Open AccessPurification, crystallization and preliminary X‐ray analysis of the strawberry allergens Fra a 1E and Fra a 3 in the presence of catechin
Author(s) -
Casañal Ana,
Zander Ulrich,
Dupeux Florine,
Valpuesta Victoriano,
Marquez Jose A.
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113006945
Subject(s) - crystallization , chemistry , escherichia coli , catechin , crystallography , biology , biochemistry , gene , polyphenol , organic chemistry , antioxidant
The strawberry Fra a proteins belong to the pathogenesis‐related PR‐10 protein family and share a common fold with the Bet v 1 major pollen allergen and the START/PYR/PYL proteins, which are characterized by the presence of a central cavity and are often involved in the binding of a variety of natural compounds. The Fra a proteins play a key role in the control of flavonoid biosynthesis in strawberries and are essential for pigment formation in fruits. In order to understand Fra a protein function, full‐length Fra a 1E and Fra a 3 cDNAs were cloned and expressed in Escherichia coli , and the proteins were purified to homogeneity using metal‐affinity chromatography. Diffraction‐quality crystals of Fra a 1E and of Fra a 3 in the presence of (+)‐catechin were obtained by the sitting‐drop vapour‐diffusion method. X‐ray diffraction data from single crystals of Fra a 1E and Fra a 3 were processed to 2.2 and 3.0 Å resolution in space groups P 2 1 2 1 2 1 and P 222 1 , with unit‐cell parameters a = 70.02, b = 74.42, c = 84.04 Å and a = 137.91, b = 206.61, c = 174.7 Å for Fra a 1E and Fra a 3, respectively.