Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay
文献类型：期刊论文
| 作者 | Ding, FX; Xu, YF; Azzi, A; Zhu, DW; Rehse, P; Chen, CQ; Sun, SH; Lin, SX |
| 刊名 | CRYSTAL GROWTH & DESIGN
 |
| 出版日期 | 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 |
| 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. |
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