A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies
A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies
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DOI：
10.1007/s10858-011-9513-5
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发表时间：
2011-07-01
影响因子：
2.7
通讯作者：
Boisbouvier, Jerome
中科院分区：
文献类型：
--
作者：
Amero, Carlos;Dura, M. Asuncion;Boisbouvier, Jerome
获得序列特异性指派仍然是溶液核磁共振研究超分子结构、动力学和相互作用的主要瓶颈。本研究表明，结合快速核磁共振实验、残基类型特异性同位素标记和自动位点定向诱变，可以有效地实现高分子量蛋白质组件中甲基探针的共振分配。通过表征涉及468 kda多聚氨基肽酶PhTET2的分子间相互作用，证明了这种通用而直接的策略的效用。
Obtaining sequence-specific assignments remains a major bottleneck in solution NMR investigations of supramolecular structure, dynamics and interactions. Here we demonstrate that resonance assignment of methyl probes in high molecular weight protein assemblies can be efficiently achieved by combining fast NMR experiments, residue-type-specific isotope-labeling and automated site-directed mutagenesis. The utility of this general and straightforward strategy is demonstrated through the characterization of intermolecular interactions involving a 468-kDa multimeric aminopeptidase, PhTET2.