Membrane domain structures of three classes of histidine kinase receptors by cell-free expression and rapid NMR analysis.

ABSTRACT: NMR structural studies of membrane proteins (MP) are hampered by complications in MP expression, technical difficulties associated with the slow process of NMR spectral peak assignment, and limited distance information obtainable for transmembrane (TM) helices. To overcome the inherent challenges in the determination of MP structures, we have developed a rapid and cost-efficient strategy that combines cell-free (CF) protein synthesis, optimized combinatorial dual-isotope labeling for nearly instant resonance assignment, and fast acquisition of long-distance information using paramagnetic probes. Here we report three backbone structures for the TM domains of the three classes of Escherichia coli histidine kinase receptors (HKRs). The ArcB and QseC TM domains are both two-helical motifs, whe

SUBMITTER: Maslennikov I 

PROVIDER: S-EPMC2890740 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature