Characterization of intrinsically disordered proteins (IDPs) has grown tremendously over the past two decades. NMR-based structural characterization has been widely embraced by the IDP community, largely because this technique is amenable to highly flexible biomolecules. Particularly, carbon-detect nuclear magnetic resonance (NMR) experiments provide a straight forward and expedient method for completing backbone assignments, thus providing the framework to study the structural and dynamic properties of IDPs. However, these experiments remain unfamiliar to most NMR spectroscopists, thus limiting the breadth of their application. In an effort to remove barriers that may prevent the application of carbon-detected bio-NMR where it has the potential to benefit investigators, here we describe the experimental requirements to collect a robust set of carbon-detected NMR data for complete backbone assignment of IDPs. Specifically, we advocate the use of three-dimensional experiments that exploit magnetization transfer pathways initiated on the aliphatic protons, which produces increased sensitivity and provides a suitable method for IDPs that are only soluble in basic pH conditions (>7.5). The applicability of this strategy to systems featuring a high degree of proline content will also be discussed.
|Original language||English (US)|
|Number of pages||13|
|Journal||Concepts in Magnetic Resonance Part A: Bridging Education and Research|
|State||Published - Jan 1 2015|
All Science Journal Classification (ASJC) codes