Open AccessA primer for carbon‐detected NMR applications to intrinsically disordered proteins in solution
Author(s) -
Bastidas Monique,
Gibbs Eric B.,
Sahu Debashish,
Showalter Scott A.
Publication year - 2015
Publication title -
concepts in magnetic resonance part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.229
H-Index - 49
eISSN - 1552-5023
pISSN - 1546-6086
DOI - 10.1002/cmr.a.21327
Subject(s) - intrinsically disordered proteins , characterization (materials science) , biomolecule , limiting , magnetization transfer , carbon fibers , materials science , chemistry , computer science , chemical physics , nuclear magnetic resonance , nanotechnology , physics , magnetic resonance imaging , engineering , algorithm , mechanical engineering , medicine , composite number , radiology
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. © 2015 Wiley Periodicals, Inc. Concepts Magn Reson Part A 43A: 54–66, 2015.