How to Prepare Membrane Proteins for Solid-State NMR: A Case Study on the α-Helical Integral Membrane Protein Diacylglycerol Kinase from E. coli

ChemBioChem ◽  
2005 ◽  
Vol 6 (9) ◽  
pp. 1693-1700 ◽  
Author(s):  
Mark Lorch ◽  
Salem Faham ◽  
Christoph Kaiser ◽  
Ingrid Weber ◽  
A. James Mason ◽  
...  
Keyword(s):  
Solid State ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Solid State Nmr ◽  
Diacylglycerol Kinase ◽  
Integral Membrane Protein ◽  
E Coli

scholarly journals Membrane Protein Structural Validation by Oriented Sample Solid-State NMR: Diacylglycerol Kinase

2014 ◽  
Vol 106 (8) ◽  
pp. 1559-1569 ◽  
Author(s):  
Dylan T. Murray ◽  
Conggang Li ◽  
F. Philip Gao ◽  
Huajun Qin ◽  
Timothy A. Cross
Keyword(s):  
Solid State ◽  
Membrane Protein ◽  
Solid State Nmr ◽  
Diacylglycerol Kinase ◽  
Oriented Sample ◽  
Structural Validation

scholarly journals Membrane Protein Structure Determination and Characterisation by Solution and Solid-State NMR

Biology ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 396
Author(s):  
Vivien Yeh ◽  
Alice Goode ◽  
Boyan B. Bonev
Keyword(s):  
Protein Structure ◽  
Solid State ◽  
High Resolution ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Solid State Nmr ◽  
Basic Unit ◽  
Molecular Assemblies ◽  
Nmr Studies ◽  
Main Challenge

Biological membranes define the interface of life and its basic unit, the cell. Membrane proteins play key roles in membrane functions, yet their structure and mechanisms remain poorly understood. Breakthroughs in crystallography and electron microscopy have invigorated structural analysis while failing to characterise key functional interactions with lipids, small molecules and membrane modulators, as well as their conformational polymorphism and dynamics. NMR is uniquely suited to resolving atomic environments within complex molecular assemblies and reporting on membrane organisation, protein structure, lipid and polysaccharide composition, conformational variations and molecular interactions. The main challenge in membrane protein studies at the atomic level remains the need for a membrane environment to support their fold. NMR studies in membrane mimetics and membranes of increasing complexity offer close to native environments for structural and molecular studies of membrane proteins. Solution NMR inherits high resolution from small molecule analysis, providing insights from detergent solubilised proteins and small molecular assemblies. Solid-state NMR achieves high resolution in membrane samples through fast sample spinning or sample alignment. Recent developments in dynamic nuclear polarisation NMR allow signal enhancement by orders of magnitude opening new opportunities for expanding the applications of NMR to studies of native membranes and whole cells.

scholarly journals Water Accessibility Refinement of the Extended Structure of KirBac1.1 in the Closed State

2021 ◽  
Vol 8 ◽  
Author(s):  
Reza Amani ◽  
Charles D. Schwieters ◽  
Collin G. Borcik ◽  
Isaac R. Eason ◽  
Ruixian Han ◽  
...  
Keyword(s):  
Solid State ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Solid State Nmr ◽  
Structural Quality ◽  
Nmr Structures ◽  
Kir Channel ◽  
C Terminus ◽  
Water Accessibility ◽  
Distance Restraints

NMR structures of membrane proteins are often hampered by poor chemical shift dispersion and internal dynamics which limit resolved distance restraints. However, the ordering and topology of these systems can be defined with site-specific water or lipid proximity. Membrane protein water accessibility surface area is often investigated as a topological function via solid-state NMR. Here we leverage water-edited solid-state NMR measurements in simulated annealing calculations to refine a membrane protein structure. This is demonstrated on the inward rectifier K+ channel KirBac1.1 found in Burkholderia pseudomallei. KirBac1.1 is homologous to human Kir channels, sharing a nearly identical fold. Like many existing Kir channel crystal structures, the 1p7b crystal structure is incomplete, missing 85 out of 333 residues, including the N-terminus and C-terminus. We measure solid-state NMR water proximity information and use this for refinement of KirBac1.1 using the Xplor-NIH structure determination program. Along with predicted dihedral angles and sparse intra- and inter-subunit distances, we refined the residues 1–300 to atomic resolution. All structural quality metrics indicate these restraints are a powerful way forward to solve high quality structures of membrane proteins using NMR.

Solid-State NMR Spectroscopic Studies of an Integral Membrane Protein Inserted into Aligned Phospholipid Bilayer Nanotube Arrays

2004 ◽  
Vol 126 (31) ◽  
pp. 9504-9505 ◽  
Author(s):  
Gary A. Lorigan ◽  
Paresh C. Dave ◽  
Elvis K. Tiburu ◽  
Krishnan Damodaran ◽  
Shadi Abu-Baker ◽  
...  
Keyword(s):  
Solid State ◽  
Membrane Protein ◽  
Solid State Nmr ◽  
Integral Membrane Protein ◽  
Spectroscopic Studies ◽  
Phospholipid Bilayer ◽  
Nanotube Arrays

Magic-angle spinning solid-state NMR of a 144 kDa membrane protein complex: E. coli cytochrome bo3 oxidase

2006 ◽  
Vol 36 (1) ◽  
pp. 55-71 ◽  
Author(s):  
Heather L. Frericks ◽  
Donghua H. Zhou ◽  
Lai Lai Yap ◽  
Robert B. Gennis ◽  
Chad M. Rienstra
Keyword(s):  
Solid State ◽  
Membrane Protein ◽  
Protein Complex ◽  
Solid State Nmr ◽  
Magic Angle Spinning ◽  
Magic Angle ◽  
Membrane Protein Complex ◽  
E Coli ◽  
Cytochrome Bo3 ◽  
Angle Spinning
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