Determination of Protein Backbone Structures from Residual Dipolar Couplings
There are a number of circumstances in which a focus on determination of the backbone structure of a protein, as opposed to a complete all-atom structure, may be appropriate. This is particularly the case for structures determined as a part of a structural genomics initiative in which computational modeling of many sequentially related structures from the backbone of a single family representative is anticipated. It is, however, also the case when the backbone may be a stepping-stone to more targeted studies of ligand interaction or protein–protein interaction. Here an NMR protocol is described that can produce a backbone structure of a protein without the need for extensive experiments directed at side chain resonance assignment or the collection of structural information on side chains. The procedure relies primarily on orientational constraints from residual dipolar couplings as opposed to distance constraints from NOEs. Procedures for sample preparation, data acquisition, and data analysis are described, along with examples from application to small target proteins of a structural genomics project.
Published in Methods in Enzymology, Volume 394, Winter 2005, pages 175-209.
© Methods in Enzymology 2005, Elseiver
Prestegard, J., Mayer, K., Valafar, H., & Benison, G. (2005). Determination of Protein Backbone Structures from Residual Dipolar Couplings. Methods In Enzymology, 175-209. doi: 10.1016/s0076-6879(05)94007-x