Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay

	Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay
	Ding, FX; Xu, YF; Azzi, A; Zhu, DW; Rehse, P; Chen, CQ; Sun, SH; Lin, SX
刊名	CRYSTAL GROWTH & DESIGN
	2010
卷号	10 期号:6 页码:2528-2532
通讯作者	Sun, SH (reprint author), Second Mil Med Univ, Dept Med Genet, Xiangyin Rd 800, Shanghai 200433, Peoples R China.,shsun@vip.sina.com ; structuralbiology@sibs.ac.cn
英文摘要	Annexin B1 (AnxB1) is a calcium-dependent phospholipid binding protein from Taenia solium cysticercus and has been reported to possess anticoagulant activity, to inhibit phospholipase A(2), and to regulate membrane transport. Native AnxB1 and its selenomethionyl derivative have been overproduced in Escherichia coli and purified. The results of dynamic light scattering analysis showed that Hepes buffer combined with low concentration salts (NaCl or CaCl2) was beneficial for preventing aggregation and for AnxB1 stabilization in the storage. After the additive screen, crystals have been yielded in the presence of guanidine hydrochloride (Gn-HCl). We determined that a low concentration of Gn-HCl significantly delayed clotting time and increased anticoagulant activity. Analysis of the crystal showed that in the presence of Gn-HCI, AnxB1 crystallizes in orthorhombic space group, which is modified from the cubic space group for crystals grown in the absence of Gn-HCl. A high quality data set (at 1.9 angstrom) has been collected successfully for crystals of L-selenomethionine labeled protein in the presence of Gn-HCl, to solve the structure with the single anomalous dispersion method (SAD). The unit cell parameters are a = 102.35 angstrom, b = 103.59 angstrom, c = 114.60 angstrom, alpha = beta = gamma = 90.00 degrees.
学科主题	Chemistry; Crystallography; Materials Science
类目[WOS]	Chemistry, Multidisciplinary ; Crystallography ; Materials Science, Multidisciplinary
关键词[WOS]	BIOCHEMICAL-CHARACTERIZATION ; BINDING PROTEIN
收录类别	SCI
语种	英语
WOS记录号	WOS:000278147900017
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1017]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Ding, FX,Xu, YF,Azzi, A,et al. Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay[J]. CRYSTAL GROWTH & DESIGN,2010,10(6):2528-2532.
APA	Ding, FX.,Xu, YF.,Azzi, A.,Zhu, DW.,Rehse, P.,...&Lin, SX.(2010).Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay.CRYSTAL GROWTH & DESIGN,10(6),2528-2532.
MLA	Ding, FX,et al."Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay".CRYSTAL GROWTH & DESIGN 10.6(2010):2528-2532.