Agriculture and Agri-Food Canada , Saskatoon, Saskatchewan, S7N 0X2 Canada.
J Agric Food Chem. 2013 Jun 19;61(24):5848-59. doi: 10.1021/jf400559x. Epub 2013 Jun 5.
Heteromeric cruciferin from wild type (WT) Arabidopsis thaliana and homomeric cruciferin CRUA, CRUB, and CRUC composed of identical subunits obtained from double-knockout mutant lines were investigated for their structural and physicochemical properties. A three-step chromatographic procedure allowed isolation of intact cruciferin hexamers with high purity (>95%). FT-IR and CD analysis of protein secondary structure composition revealed that all cruciferins were folded into higher order structures consisting of 44-50% β-sheets and 7-9% α-helices. The structural and physicochemical properties of homohexameric CRUC deviated from that of CRUA and CRUB and exhibited a compact, thermostable, and less hydrophobic structure, confirming the predictions made using 3D homology structure models.
野生型拟南芥异源六聚体芸薹素和同源六聚体 CRUA、CRUB、CRUC 是由双敲除突变系获得的相同亚基组成，研究了它们的结构和物理化学性质。三步色谱分离程序可实现高纯度 (>95%)的完整芸薹素六聚体的分离。蛋白质二级结构组成的 FT-IR 和 CD 分析表明，所有芸薹素均折叠成由 44-50%β-折叠和 7-9%α-螺旋组成的高级结构。同六聚体 CRUC 的结构和物理化学性质与 CRUA 和 CRUB 不同，表现出紧凑、热稳定和疏水性较小的结构，证实了使用 3D 同源结构模型的预测。
