[NMR paper] Multidimensional Solid-State NMR of a Functional Multiprotein Chemoreceptor Array.

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Multidimensional Solid-State NMR of a Functional Multiprotein Chemoreceptor Array.

Related Articles Multidimensional Solid-State NMR of a Functional Multiprotein Chemoreceptor Array.

Biochemistry. 2016 Jun 13;

Authors: Harris MJ, Struppe JO, Wylie BJ, McDermott AE, Thompson LK

Abstract
The bacterial chemoreceptor complex governs signal detection and the upstream elements of chemotactic behavior, but the detailed molecular mechanism is still unclear. We have assembled native-like functional arrays of an aspartate receptor cytoplasmic fragment (CF) with its two cytoplasmic protein partners (CheA and CheW) for solid-state NMR studies of structural changes involved in signaling. In this initial study of the uniformly 13C,15N-enriched CF in these >13.8 MDa size arrays, residue-type assignments are made for amino acids that together make up 90% of the protein. We demonstrate that homo and heteronuclear two-dimensional spectra are consistent with structure-based chemical shift predictions: a number of major assignable correlations are consistent with the predominantly alpha helical secondary structure and minor correlations are consistent with the disordered C-terminal tail. Sub-ppm linewidths and spectral changes on sample freezing suggest these arrays are structurally homogeneous and sufficiently immobilized for efficient solid-state NMR.


PMID: 27295350 [PubMed - as supplied by publisher]



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