Unambiguous Determination of Protein Arginine Ionization States in Solution by NMR SpectroscopyYoshimura, Y., Oktaviani, N. A., Yonezawa, K., Kamikubo, H. & Mulder, F. A. A., 2-Jan-2017, In : Angewandte Chemie-International Edition. 56, 1, p. 239-242 4 p.
Research output: Contribution to journal › Article › Academic › peer-review
Because arginine residues in proteins are expected to be in their protonated form almost without exception, reports demonstrating that a protein arginine residue is charge-neutral are rare and potentially controversial. Herein, we present a C-13-detected NMR experiment for probing individual arginine residues in proteins notwithstanding the presence of chemical and conformational exchange effects. In the experiment, the N-15(eta) and N-15(epsilon) chemical shifts of an arginine head group are correlated with that of the directly attached C-13(zeta). In the resulting spectrum, the number of protons in the arginine head group can be obtained directly from the N-15-H-1 scalar coupling splitting pattern. We applied this method to unambiguously determine the ionization state of the R52 side chain in the photoactive yellow protein from Halorhodospira halophila. Although only three H-eta atoms were previously identified by neutron crystallography, we show that R52 is predominantly protonated in solution.
|Number of pages||4|
|Journal||Angewandte Chemie-International Edition|
|Publication status||Published - 2-Jan-2017|
- arginine, cross polarization, N-15-H-1 spin-spin coupling, NMR spectroscopy, photoactive yellow protein, PHOTOACTIVE YELLOW PROTEIN, INTRINSICALLY DISORDERED PROTEINS, SIDE-CHAIN DYNAMICS, HYDROGEN-BONDS, ACTIVE-SITE, N-15, RESIDUES, LYSINE, BACTERIORHODOPSIN, THERMOSTABILITY