A device for the measurement of residual chemical shift anisotropy and residual dipolar coupling in soluble and membrane-associated proteins.
Residual dipolar coupling (RDC) and residual chemical shift anisotropy (RCSA) report on orientational properties of a dipolar bond vector and a chemical shift anisotropy principal axis system, respectively. They can be highly complementary in the analysis of backbone structure and dynamics in proteins as RCSAs generally include a report on vectors out of a peptide plane while RDCs usually report on in-plane vectors. Both RDC and RCSA average to zero in isotropic solutions and require partial orientation in a magnetic field to become observable. While the alignment and measurement of RDC has become routine, that of RCSA is less common. This is partly due to difficulties in providing a suitable isotopic reference spectrum for the measurement of the small chemical shift offsets coming from RCSA. Here we introduce a device (modified NMR tube) specifically designed for accurate measurement of reference and aligned spectra for RCSA measurements, but with a capacity for RDC measurements as well. Applications to both soluble and membrane anchored proteins are illustrated.
Liu
Yizhou
Y
Complex Carbohydrate Research Center, The University of Georgia, Athens, GA 30602, USA.
Prestegard
James H
JH
D000180:Acrylic Resins / Q000737:chemistry; D000465:Algorithms; D016880:Anisotropy; D016018:Least-Squares Analysis; D008565:Membrane Proteins / Q000737:chemistry*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000295:instrumentation* / Q000379:methods; D011506:Proteins / Q000737:chemistry*; D000099166:Polyacrylamides
D000180:Acrylic Resins; D008565:Membrane Proteins; D011506:Proteins; D000099166:Polyacrylamides