化学交联和 LC/MS 分析确定蛋白质结构域取向：在 AbrB 中的应用。
Chemical crosslinking and LC/MS analysis to determine protein domain orientation: application to AbrB.
机构信息
Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695, USA.
出版信息
Biochem Biophys Res Commun. 2013 Feb 8;431(2):253-7. doi: 10.1016/j.bbrc.2012.12.124. Epub 2013 Jan 9.
Abstract
To fully understand the modes of action of multi-protein complexes, it is essential to determine their overall global architecture and the specific relationships between domains and subunits. The transcription factor AbrB is a functional homotetramer consisting of two domains per monomer. Obtaining the high-resolution structure of tetrameric AbrB has been extremely challenging due to the independent character of these domains. To facilitate the structure determination process, we solved the NMR structures of both domains independently and utilized gas-phase cleavable chemical crosslinking and LC/MS(n) analysis to correctly position the domains within the full tetrameric AbrB protein structure.
摘要
为了充分了解多蛋白复合物的作用模式，确定其整体全局结构以及结构域和亚基之间的特定关系至关重要。转录因子 AbrB 是由每个单体的两个结构域组成的功能性同源四聚体。由于这些结构域具有独立性，因此获得四聚体 AbrB 的高分辨率结构极具挑战性。为了促进结构确定过程，我们分别独立地解决了两个结构域的 NMR 结构，并利用气相可裂解化学交联和 LC/MS(n) 分析，正确地将结构域定位在完整的四聚体 AbrB 蛋白质结构内。
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