Structural Characterization of a Flexible Two-Domain Protein in Solution Using Small Angle X-Ray Scattering and NMR Data
 

Structural Characterization of a Flexible Two-Domain Protein in Solution Using Small Angle X-Ray Scattering and NMR Data

Publication date: Available online 6 November 2014
Source:Structure</br>
Author(s): Alexander Lemak , Bin Wu , Adelinda Yee , Scott Houliston , Hsiau-Wei Lee , Aleksandras Gutmanas , Xianyang Fang , Maite Garcia , Anthony Semesi , Yun-Xing Wang , James*H. Prestegard , Cheryl*H. Arrowsmith</br>
Multidomain proteins in which individual domains are connected by linkers often possess inherent interdomain flexibility that significantly complicates their structural characterization in solution using either nuclear magnetic resonance (NMR) spectroscopy or small-angle X-ray scattering (SAXS) alone. Here, we report a protocol for joint refinement of flexible multidomain protein structures against NMR distance and angular restraints, residual dipolar couplings, and SAXS data. The protocol is based on the ensemble optimization method principle (Bernadó et*al., 2007) and is compared with different refinement strategies for the structural characterization of the flexible two-domain protein sf3636 from Shigella flexneri 2a. The results of our refinement suggest the existence of a dominant population of configurational states in solution possessing an overall elongated shape and restricted relative twisting of the two domains.
</br> Teaser

Lemak et*al. describe a protocol for joint ensemble refinement of flexible multidomain protein structures against NOE, RDC, and SAXS data and apply it to a two-domain protein sf3636 from Shigella flexneri 2a. Sf3636 populates structural states with an extended arrangement of two domains that display a restricted twisting motion.</br>
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