Novel techniques for weak alignment of proteins in solution using chemical tags coordinating lanthanide ions.
A molecule with an anisotropic magnetic susceptibility is spontaneously aligned in a static magnetic field. Alignment of such a molecule yields residual dipolar couplings and pseudocontact shifts. Lanthanide ions have recently been successfully used to provide an anisotropic magnetic susceptibility in target molecules either by replacing a calcium ion with a lanthanide ion in calcium-binding proteins or by attaching an EDTA derivative to a cysteine residue via a disulfide bond. Here we describe a novel enantiomerically pure EDTA derived tag that aligns stronger due to its shorter linker and does not suffer from stereochemical diversity upon lanthanide complexation. We observed residual (15)N,(1)H-dipolar couplings of up to 8 Hz at 800 MHz induced by a single alignment tensor from this tag.
Ikegami
Takahisa
T
Max-Planck Institute for Biophysical Chemistry, Department of NMR Based Structural Biology, University of Frankfurt am Fassberg 11, D-37077 Göttingen, Germany.
Verdier
Laurent
L
Sakhaii
Peyman
P
Grimme
Susanne
S
Pescatore
Barbara
B
Saxena
Krishna
K
Fiebig
Klaus M
KM
Griesinger
Christian
C
D016880:Anisotropy; D002118:Calcium / Q000737:chemistry; D002255:Carbonic Acid / Q000737:chemistry; D004220:Disulfides; D004492:Edetic Acid / Q000737:chemistry; D007477:Ions; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008956:Models, Chemical; D008958:Models, Molecular; D015233:Models, Statistical; D009584:Nitrogen; D011485:Protein Binding; D011487:Protein Conformation; D013237:Stereoisomerism
D004220:Disulfides; D007477:Ions; D028581:Lanthanoid Series Elements; D002255:Carbonic Acid; D004492:Edetic Acid; D009584:Nitrogen; D002118:Calcium