10 Feb 2021
10 Feb 2021
Real-time NMR spectroscopy in the study of biomolecular kinetics and dynamics
- 1Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, 60438, Germany
- 2Institute for Biophysical Chemistry, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, 60438, Germany
- 1Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, 60438, Germany
- 2Institute for Biophysical Chemistry, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, 60438, Germany
Abstract. The review describes the application of NMR spectroscopy to study kinetics of folding, refolding and aggregation of proteins, RNA and DNA. Time-resolved NMR experiments can be conducted in a reversible or an irreversible manner. In particular irreversible folding experiments pose large requirements on (i) the signal-to-noise due to the time limitations and (ii) on synchronizing the refolding steps. Thus, this contribution discusses the application of methods for signal-to-noise increases including dynamic nuclear polarization, hyperpolarization and photo-CIDNP for the study of time-resolved NMR studies. Further, methods are reviewed ranging from pressure- and temperature-jump, light induction and rapid mixing to induce rapidly non-equilibrium conditions required to initiate folding.
György Pintér et al.
Status: open (until 21 Mar 2021)
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RC1: 'Comment on mr-2021-16', Peter Hore, 15 Feb 2021
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An excellent review and an enjoyable read. A fitting tribute to Rob.
I spotted a trivial number of minor inaccuracies and spelling errors.
Lines 50, 53, 55: by describing photo-CIDNP dyes as “fluorescent” and “fluorophores” you give the impression that fluorescence if somehow essential for the generation of nuclear polarization. Efficient fluorescence in competition with intersystem crossing would, of course, be a distinct disadvantage in sensitivity terms.
Line 54: did you mean to include phenylalanine amongst the list of polarizable amino acids?
Line 183: photoactive yellow protein?
Line 187: shouldn’t free enthalpy be simply enthalpy or enthalpy change?
Line 278: dilution factor?
Lines 283-284: incomplete sentence?
Lines 304 and 330: CIDNP not CINDP
Lines 306-307: “influenced by the hyperfine coupling constants of the present magnetic field” makes no sense.
Line 361: irradiation with argon-ion laser light?
Line 456: channel rhodopsin
Line 548: nitrobenzyl
Line 670: longitudinal proton relaxation time?
Not all abbreviations are defined (or used more than once). While I think this is acceptable for NMR pulse sequence acronyms, there are others which will be less familiar to readers of this review, for example: FAD, FMN, GPCR, TFE, DMSO, CSNB, D+PHS, RAS, TET2, …
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AC1: 'Reply on RC1', Harald Schwalbe, 15 Feb 2021
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Dear Peter,
thank you for your nice comments. We will of course correct the points in the final version of the manuscript.
We will wait before we do so to see additional comments.
best wishes
H. Schwalbe
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AC1: 'Reply on RC1', Harald Schwalbe, 15 Feb 2021
reply
György Pintér et al.
György Pintér et al.
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