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Unambiguous Determination of Protein Arginine Ionization States in Solution by NMR Spectroscopy

Yoshimura, 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.

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  • Unambiguous Determination of Protein Arginine Ionization States in Solution

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DOI

  • Yuichi Yoshimura
  • Nur Alia Oktaviani
  • Kento Yonezawa
  • Hironari Kamikubo
  • Frans A. A. Mulder

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.

Original languageEnglish
Pages (from-to)239-242
Number of pages4
JournalAngewandte Chemie-International Edition
Volume56
Issue number1
Publication statusPublished - 2-Jan-2017

    Keywords

  • 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

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