Details

Autor(en) / Beteiligte

• Sonntag, Miriam
• Jagtap, Pravin Kumar Ankush
• Simon, Bernd
• Appavou, Marie‐Sousai
• Geerlof, Arie
• Stehle, Ralf
• Gabel, Frank
• Hennig, Janosch
• Sattler, Michael

Titel

Segmental, Domain‐Selective Perdeuteration and Small‐Angle Neutron Scattering for Structural Analysis of Multi‐Domain Proteins

Ist Teil von

• Angewandte Chemie International Edition, 2017-08, Vol.56 (32), p.9322-9325

Auflage

International ed. in English

Ort / Verlag

Germany: Wiley Subscription Services, Inc

Erscheinungsjahr

2017

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. Segmentally perdeuterated multi‐domain proteins are obtained by Sortase A mediated ligation. Using contrast‐matched small‐angle neutron scattering (SANS) experiments on different samples with selectively perdeuterated domains of the same multi‐domain protein gives insight into domain arrangements and changes induced by ligand binding, thus providing unique information for structural analysis.