High level expression and dimer characterization of the S100 EF-hand proteins, migration inhibitory factor-related proteins 8 and 14
High level expression and dimer characterization of the S100 EF-hand proteins, migration inhibitory factor-related proteins 8 and 14
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DOI：
10.1074/jbc.273.20.12427
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发表时间：
1998-05-15
影响因子：
4.8
通讯作者：
Chazin, WJ
中科院分区：
文献类型：
--
作者：
Hunter, MJ;Chazin, WJ
不同巨噬细胞亚群的表型和功能异质性由两种S100钙结合蛋白（迁移抑制因子相关蛋白（MRP）8和14）表达的离散变化定义。为了进一步了解MRP 8和MRP 14在炎症反应的发展阶段，通过基于T7的表达载体和大肠杆菌BL 21（DE 3）细胞系的组合获得MRPs的过表达。一个有效的，两步色谱协议，然后开发快速，简便的纯化。广泛的生物物理表征和化学交联实验表明，MRP 8和MRP 14形成的寡聚体具有强烈的偏好，以缔合为异二聚体。异源NMR实验表明，一个特定的良好包装的二聚体只形成在两种蛋白质的等摩尔混合物。我们的研究结果表明，有一个独特的互补性，在界面的MRP 8/MRP 14复合物，不能完全复制的MRP 8和MRP 14同源二聚体。
The phenotypical and functional heterogeneity of different macrophage subpopulations are defined by discrete changes in the expression of two S100 calcium-binding proteins, migration inhibitory factor-related proteins (MRPs) 8 and 14. To further our understanding of MRP8 and MRP14 in the developmental stages of inflammatory responses, overexpression of the MRPs was obtained through a combination of a T7-based expression vector and the Escherichia coli BL21 (DE3) cell line. An efficient, two-step chromatographic protocol was then developed for rapid, facile purification. Extensive biophysical characterization and chemical cross linking experiments show that MRP8 and MRP14 form oligomers with a strong preference to associate as a heterodimer. Heteronuclear NMR experiments indicate that a specific well packed dimer is formed only in equimolar mixtures of the two proteins. Our results suggest that there is a unique complementarity in the interface of the MRP8/MRP14 complex that cannot be fully reproduced in the MRP8 and MRP14 homodimers.