NMR paper Automated backbone NMR resonance assignment of large proteins using redundant linking from a single simultaneous acquisition.

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[NMR paper] Automated backbone NMR resonance assignment of large proteins using redundant linking from a single simultaneous acquisition.

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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

Quality Control Check

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

From NMR-STAR 3.1

Validate NMR-STAR 3.1

NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

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03-07-2020, 06:20 PM

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Automated backbone NMR resonance assignment of large proteins using redundant linking from a single simultaneous acquisition.

Related Articles Automated backbone NMR resonance assignment of large proteins using redundant linking from a single simultaneous acquisition.

J Am Chem Soc. 2020 Mar 04;:

Authors: Stanek J, Schubeis T, Paluch P, Güntert P, Andreas LB, Pintacuda G

Abstract
Thanks to magic-angle spinning (MAS) probes with frequencies*of 60-100 kHz, the benefit of high sensitivity*1H detection can now*be broadly realized in biomolecular solid-state NMR for the analysis of*microcrystalline,*sedimented, or lipid-embedded preparations. Nonetheless, performing*the assignment of all resonances remains*a*rate-limiting step in protein*structural studies, and even the latest optimized protocols fail to perform this*step when the protein size exceeds ~20 kDa. Here we leverage the benefits of*fast (100 kHz) MAS and high (800 MHz) magnetic fields to design an approach that*lifts this limitation. Through the creation, conservation and*acquisition of*independent magnetization pathways within a single triple-resonance MAS NMR*experiment, a single self-consistent dataset can be acquired, providing enhanced sensitivity, reduced vulnerability to*machine or sample instabilities,*and highly redundant linking that supports fully-automated peak picking and resonance assignment. The method, dubbed RAVASSA (Redundant Assignment Via A Single*Simultaneous Acquisition), is demonstrated with the assignment of the largest protein*to date in the solid state, the 42.5 kDa maltose binding protein, using a single fully protonated*microcrystalline sample and one week of spectrometer time.

PMID: 32129995 [PubMed - as supplied by publisher]

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