NMR paper Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy.

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[NMR paper] Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy.

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NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

Preditor

TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

Flexibility from chemical shifts:

RCI

Interactions from chemical shifts:

HADDOCK

Chemical shifts re-referencing:

NMR model quality:

NOEs, other restraints:

Chemical shifts:

RDCs:

Pseudocontact shifts:

Protein geomtery:

SAVES2 or SAVES4

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NMR spectrum prediction:

FANDAS

MestReS

V-NMR

Flexibility from structure:

Backbone S2

Methyl S2

B-factor

Molecular dynamics:

Gromacs

Amber

Antechamber

Chemical shifts prediction:

From structure:

Shiftx2

Sparta+

Camshift

CH3shift- Methyl

ArShift- Aromatic

ShiftS

Proshift

PPM

CheShift-2- Cα

From sequence:

Shifty

Camcoil

Poulsen_rc_CS

Disordered proteins:

MAXOCC

Format conversion & validation:

CCPN

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NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

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08-27-2015, 12:37 PM

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Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy.

Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy.

Related Articles Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy.

Anal Chem. 2015 Aug 26;

Authors: Bugge K, Steinocher H, Brooks AJ, Lindorff-Larsen K, Kragelund BB

Abstract
Despite the biological and pharmaceutical significance of membrane proteins, their tertiary structures constitute less than 3% of known structures. One of the major obstacles for initiating structural studies of membrane proteins by NMR spectroscopy is the generation of high amounts of isotope-labeled protein. In this work we have exploited the hydrophobic nature of membrane proteins to develop a simple and efficient production scheme for isotope-labeled single-pass transmembrane domains (TMDs) with or without intrinsically disordered regions. We have evaluated the applicability and limitations of the strategy using seven membrane protein variants that differ in their overall hydrophobicity and length, and show a recovery for suitable variants of >70%. The developed production scheme is cost-efficient and easy to implement, and has the potential to facilitate an increase in the number of structures of single-pass TMDs, which are difficult to solve by other means.

PMID: 26309151 [PubMed - as supplied by publisher]

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