NMR and small-angle scattering-based structural analysis of protein complexes in solution

NMR Spectroscopy for Studying Interactions of Bioactive Molecules

“…Madl et al [111] discussed the utility and computational approaches of combining solution-state NMR with small-angle X-ray and neutron scattering (SAXS/SANS) experiments for the structural analysis of large protein complexes. The combination of NMR and SAXS/SANS in particular is a valuable approach that complements crystallographic studies for structure determination and the investigation of dynamic properties of multi-domain proteins and their complexes in solution.…”

EPR-aided approach for solution structure determination of large RNAs or protein–RNA complexes

“…The measurement of SAXS data has proved to be very valuable to obtain information on the global shape of macromolecules and their complexes 8,11,43 . However, SAXS data only provides low resolution information and has to be supplemented with data obtained by complementary biophysical methods such as PREs or RDCs.…”

“…In such systems, the EPR constraints could be supplemented by additional NMR-based restraints, such as RDC or PRE data or data from other biophysical methods like SAXS [7][8][9]11 . This would reduce the number of EPR distances to be measured to obtain a unique solution of high precision.…”

“…Efficient approaches for solution structure determination of large proteins and protein-protein complexes have successfully been applied by combining traditional NMR spectroscopy with the measurement of paramagnetic relaxation enhancement (PRE), residual dipolar coupling (RDC) data or with other biophysical methods such as small-angle X-ray scattering (SAXS) [7][8][9][10][11] . However, these approaches fail on large RNAs and their complexes due to high-resonance overlap, line broadening and low proton density found in the RNA part of such complexes 12 .…”

Combining NMR and EPR to Determine Structures of Large RNAs and Protein–RNA Complexes in Solution

“…More recently, NMR restraints have been combined with lower resolution global shape information resulting from small angle X-ray (SAXS) and neutron scattering (SANS) measurements (Burke, Sashital, Zuo, Wang, & Butcher, 2012;Grishaev, Wu, Trewhella, & Bax, 2005;Grishaev, Ying, Canny, Pardi, & Bax, 2008;Lapinaite et al, 2013;Madl, Gabel, & Sattler, 2011) or envelops resulting from EM .…”