High-Sensitivity Rheo-NMR Spectroscopy for Protein Studies
journal contributionposted on 30.06.2017, 00:00 by Daichi Morimoto, Erik Walinda, Naoto Iwakawa, Mayu Nishizawa, Yasushi Kawata, Akihiko Yamamoto, Masahiro Shirakawa, Ulrich Scheler, Kenji Sugase
Shear stress can induce structural deformation of proteins, which might result in aggregate formation. Rheo-NMR spectroscopy has the potential to monitor structural changes in proteins under shear stress at the atomic level; however, existing Rheo-NMR methodologies have insufficient sensitivity to probe protein structure and dynamics. Here we present a simple and versatile approach to Rheo-NMR, which maximizes sensitivity by using a spectrometer equipped with a cryogenic probe. As a result, the sensitivity of the instrument ranks highest among the Rheo-NMR spectrometers reported so far. We demonstrate that the newly developed Rheo-NMR instrument can acquire high-quality relaxation data for a protein under shear stress and can trace structural changes in a protein during fibril formation in real time. The described approach will facilitate rheological studies on protein structural deformation, thereby aiding a physical understanding of shear-induced amyloid fibril formation.