In-Cell Protein Structures from 2D NMR Experiments.
In-Cell Protein Structures from 2D NMR Experiments.
J Phys Chem Lett. 2016 Jul 5;
Authors: Müntener T, Häussinger D, Selenko P, Theillet FX
Abstract
In-cell NMR spectroscopy provides atomic resolution insights into the structural properties of proteins in cells. However, it is rarely used to solve entire protein structures de novo. Here, we introduce a paramagnetic lanthanide-tag to simultaneously measure protein pseudocontact shifts (PCSs) and residual dipolar couplings (RDCs) to be used as input for structure calculation routines within the Rosetta program. We employ this approach to determine the structure of the protein G B1 domain (GB1) in intact Xenopus laevis oocytes from 1H-15N 2D in-cell NMR experiments. Specifically, we derive well-defined GB1 ensembles from low concentration in-cell NMR samples (~50 ?M) measured at moderate magnetic field strengths (600 MHz), thus offering novel routes for intracellular structure determination with data from 2D in-cell NMR experiments.
PMID: 27379949 [PubMed - as supplied by publisher]
More...