TY - JOUR
T1 - DMSO-Quenched H/D-Exchange 2D NMR Spectroscopy and Its Applications in Protein Science
AU - Kuwajima, Kunihiro
AU - Yagi-Utsumi, Maho
AU - Yanaka, Saeko
AU - Kato, Koichi
N1 - Funding Information:
Funding: This work was supported in part by JSPS (Japan Society for the Promotion of Science) KAKENHI Grant Number JP20K06574 and by Joint Research of the Exploratory Research Center on Life and Living Systems (ExCELLS) Program Number 22EXC315. A part of this work was conducted in Institute for Molecular Science, and supported by the Nanotechnology Platform Program <Molecule and Material Synthesis> (JPMXP09S21MS0021) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.
Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/6/1
Y1 - 2022/6/1
N2 - Hydrogen/deuterium (H/D) exchange combined with two-dimensional (2D) NMR spectroscopy has been widely used for studying the structure, stability, and dynamics of proteins. When we apply the H/D-exchange method to investigate non-native states of proteins such as equilibrium and kinetic folding intermediates, H/D-exchange quenching techniques are indispensable, because the exchange reaction is usually too fast to follow by 2D NMR. In this article, we will describe the dimethylsulfoxide (DMSO)-quenched H/D-exchange method and its applications in protein science. In this method, the H/D-exchange buffer is replaced by an aprotic DMSO solution, which quenches the exchange reaction. We have improved the DMSO-quenched method by using spin desalting columns, which are used for medium exchange from the H/D-exchange buffer to the DMSO solution. This improvement has allowed us to monitor the H/D exchange of proteins at a high concentration of salts or denaturants. We describe methodological details of the improved DMSO-quenched method and present a case study using the improved method on the H/D-exchange behavior of unfolded human ubiquitin in 6 M guanidinium chloride.
AB - Hydrogen/deuterium (H/D) exchange combined with two-dimensional (2D) NMR spectroscopy has been widely used for studying the structure, stability, and dynamics of proteins. When we apply the H/D-exchange method to investigate non-native states of proteins such as equilibrium and kinetic folding intermediates, H/D-exchange quenching techniques are indispensable, because the exchange reaction is usually too fast to follow by 2D NMR. In this article, we will describe the dimethylsulfoxide (DMSO)-quenched H/D-exchange method and its applications in protein science. In this method, the H/D-exchange buffer is replaced by an aprotic DMSO solution, which quenches the exchange reaction. We have improved the DMSO-quenched method by using spin desalting columns, which are used for medium exchange from the H/D-exchange buffer to the DMSO solution. This improvement has allowed us to monitor the H/D exchange of proteins at a high concentration of salts or denaturants. We describe methodological details of the improved DMSO-quenched method and present a case study using the improved method on the H/D-exchange behavior of unfolded human ubiquitin in 6 M guanidinium chloride.
UR - http://www.scopus.com/inward/record.url?scp=85131730615&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85131730615&partnerID=8YFLogxK
U2 - 10.3390/molecules27123748
DO - 10.3390/molecules27123748
M3 - Review article
C2 - 35744871
AN - SCOPUS:85131730615
SN - 1420-3049
VL - 27
JO - Molecules
JF - Molecules
IS - 12
M1 - 3748
ER -