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Sonntag, Miriam ; Jagtap, Pravin Kumar Ankush ; Simon, Bernd ; Appavou, Marie-Sousai ; Geerlof, Arie ; Stehle, Ralf ; Gabel, Frank ; Hennig, Janosch ; Sattler, Michael:
Segmental, Domain-Selective Perdeuteration and Small-Angle Neutron Scattering for Structural Analysis of Multi-Domain Proteins.
In: Angewandte Chemie International Edition.
Bd. 56
(2017)
Heft 32
.
- S. 9322-9325.
ISSN 1521-3773
DOI: https://doi.org/10.1002/anie.201702904
Abstract
Multi-domain proteins play critical roles in fine-tuning essential processes in cellular signaling and gene regulation. Typically, multiple globular domains that are connected by flexible linkers undergo dynamic rearrangements upon binding to protein, DNA or RNA ligands. RNA binding proteins (RBPs) represent an important class of multi-domain proteins, which regulate gene expression by recognizing linear or structured RNA sequence motifs. Here, we employ segmental perdeuteration of the three RNA recognition motif (RRM) domains in the RBP TIA-1 using Sortase A mediated protein ligation. We show that domain-selective perdeuteration combined with contrast-matched small-angle neutron scattering (SANS), SAXS and computational modeling provides valuable information to precisely define relative domain arrangements. The approach is generally applicable to study conformational arrangements of individual domains in multi-domain proteins and changes induced by ligand binding.