Determination of residual dipolar couplings in homonuclear MOCCA-SIAM experiments.
In solutions with partial molecular alignment, anisotropic magnetic interactions such as the chemical shift anisotropy, the electric quadrupole interaction, and the magnetic dipole-dipole interaction are no longer averaged out to zero in contrast to isotropic solutions. The resulting residual anisotropic magnetic interactions are increasingly used in biological NMR studies for the determination of 3D structures of proteins and other biomolecules. In the present paper we propose a new approach allowing the measurement of residual HN-H(alpha) dipolar couplings of non-isotope enriched proteins based on the application of the MOCCA-SIAM experiment. This experiment allows the measurement of homonuclear coupling constants with an accuracy of ca. +/- 0.2 Hz and is therefore particularly well suited to determine residual dipolar couplings at relatively low degrees of molecular orientation. The agreement between experimentally determined residual HN-H(alpha) couplings and calculated values is demonstrated for BPTI.
Möglich
Andreas
A
University of Regensburg, Institute of Biophysics and Physical Biochemistry, Germany.
Wenzler
Michael
M
Kramer
Frank
F
Glaser
Steffen J
SJ
Brunner
Eike
E
D000818:Animals; D016880:Anisotropy; D007611:Aprotinin / Q000737:chemistry; D008280:Magnetics; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D011522:Protons
D011506:Proteins; D011522:Protons; D007611:Aprotinin