Amide additives improve RDC measurements in polyacrylamide

Academic Article

Abstract

  • © 2020, Springer Nature B.V. Residual dipolar couplings (RDCs) provide valuable NMR parameters that can be used for structural calculation and verification. Measuring RDCs requires aligning macromolecules using one of various types of alignment media. Of different alignment media options, stretched or compressed polyacrylamide gels are advantageous due to their chemical stability. However, polyacrylamide interacts with proteins and significantly broadens NMR resonances. In this study, we found that the amide-containing compounds asparagine, glutamine and propionamide improve spectral quality of proteins in polyacrylamide gel without significantly reducing the magnitude of RDC values. Moreover, we showed that propionamide is an attractive additive that increases protein solubility without interfering with protein stability, ligand binding or NMR pulse width, suggesting its potential applications for our NMR methods.
  • Authors

    Published In

    Digital Object Identifier (doi)

    Author List

  • Fargason T; Wang T; De Silva NIU; Zhang Z; McKelvey H; Knapp T; Zaharias S; Zhang J
  • Start Page

  • 119
  • End Page

  • 124
  • Volume

  • 74
  • Issue

  • 2-3