Question from NMRWiki Q&A forum Internal referencing in direct and indirect dimensions using guanidinium

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[Question from NMRWiki Q&A forum] Internal referencing in direct and indirect dimensions using guanidinium

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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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11-18-2010, 06:14 AM

nmrlearner

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Internal referencing in direct and indirect dimensions using guanidinium

Internal referencing in direct and indirect dimensions using guanidinium

Hello,

I am recording 15N-HSQC spectra of a protein in increasing concentrations (0.25–2.5 M) of guanidine hydrochloride. I would like to use the guanidine chemical shift (which appears in the HSQC) as an internal standard in both the direct and indirect (!) dimensions. My intuition is that the chemical shifts of guanidine should be independent of guanidine concentration. Does anyone know if this is really true?

Currently, I see a constant upfield shift in the indirect dimension, even for the guanidine peak itself, on the order of 0.3 ppm.The pH of the samples is constant, within error. The measurements are performed at the same temperature.

Thanks in advance!

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