Open AccessPurification, crystallization and preliminary crystallographic analysis of soybean mature glycinin A1bB2
Author(s) -
Prak Krisna,
Mikami Bunzo,
Itoh Takafumi,
Fukuda Takako,
Maruyama Nobuyuki,
Utsumi Shigeru
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113019684
Subject(s) - random hexamer , ammonium sulfate , crystallization , crystallography , imidazole , chemistry , sodium citrate , ammonium phosphate , phosphate , molecule , sodium , chromatography , stereochemistry , biochemistry , fertilizer , organic chemistry , medicine , pathology
Glycinin is one of the most abundant storage‐protein molecules in soybean seeds and is composed of five subunits (A1aB1b, A1bB2, A2B1a, A3B4 and A5A4B3). A1bB2 was purified from a mutant soybean cultivar containing glycinin composed of only A5A4B3 and A1bB2. At 281 K the protein formed hexagonal, rectangular and rod‐shaped crystals in the first [0.1 M imidazole pH 8.0, 0.2 M MgCl 2 , 35%( v / v ) MPD], second [0.1 M sodium citrate pH 5.6, 0.2 M ammonium acetate, 30%( v / v ) MPD] and third (0.1 M phosphate–citrate pH 4.2, 2.0 M ammonium sulfate) crystallization conditions, respectively. X‐ray diffraction data were collected to resolutions of 1.85, 1.85 and 2.5 Å from crystals of the three different shapes. The crystals belonged to space groups P 6 3 22, P 2 1 and P 1, with unit‐cell parameters a = b = 143.60, c = 84.54 Å, a = 114.54, b = 105.82, c = 116.67 Å, β = 94.99° and a = 94.45, b = 94.96, c = 100.66 Å, α = 107.02, β = 108.44, γ = 110.71°, respectively. One, six and six subunits of A1bB2 were estimated to be present in the respective asymmetric units. The three‐dimensional structure of the A1bB2 hexamer is currently being determined.