The Inescapable Effects of Ribosomes on In-Cell NMR Spectroscopy and the Implications for Regulation of Biological Activity
Abstract
:1. Introduction
2. Protein-RNA Interactions Broaden Target Protein NMR Spectra
3. Resolving Target Protein Bound to RNA
4. Target Protein-RNA Complexes Exhibit Megadalton Apparent Molecular Masses
5. RNA-Mediated Quinary Interaction Surfaces
5.1. Adenylate Kinase
5.2. Thioredoxin
5.3. Dihydrofolate reductase and Thymidylate synthase
6. Ribosome-Mediated Regulation of Biological Activity
6.1. Adenylate Kinase
6.2. Dihydrofolate Reductase and Thymidylate Synthase
7. Discussion
Author Contributions
Funding
Conflicts of Interest
References
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Protein | Binds | Effect of RNA-Binding |
---|---|---|
Ubiquitin (Ubq) | Total RNA [14,35] mRNA [28] | Blocks polyubiquitination sites, increases apparent MW [14] |
Thioredoxin (Trx) | Total RNA [14], mRNA [28] | Increases apparent MW [14] Antibiotic binding to ribosome alters quinary structure [36] |
Adenylate kinase (ADK) | Total RNA [14] mRNA [28] Ribosome [26,29] | Increases apparent MW [14] Noncompetitive kinetic inhibitor [26] |
Dihydrofolate reductase (DHFR) | mRNA [37,38] Ribosome [26] | Competitive kinetic inhibitor [26] |
Thymidylate synthase (TS) | Total RNA [26] mRNA [28,39] Ribosome [26,29] | Uncompetitive kinetic activator [26] |
Enzyme | [ribosome] (μM) | Vmax (S−1) | VmaxRibosome/Vomax a | KM (μM) b | KI (mM) b | R2 |
---|---|---|---|---|---|---|
ADK | 0 | (4.2 ± 0.2) × 10−2 | ∼0.5 | 180 ± 20 | 6.1 ± 0.8 | 0.98 |
1 | (2.1 ± 0.1) × 10−2 | 130 ± 30 | 35 ± 2 | 0.96 | ||
DHFR | 0 | (1.48 ± 0.04) × 10−4 | ∼0.8 | 0.32 ± 0.04 | 0.95 | |
0.5 | (1.16 ± 0.02) × 10−4 | 3.5 ± 0.1 | 0.99 | |||
TS | 0 | (9.7 ± 0.4) × 10−5 | ∼20 | 5.4 ± 0.7 | 0.97 | |
0.5 | (2.0 ± 0.2) × 10−3 | 120 ± 20 | (3.9 ± 0.5) × 10−3 | 0.99 |
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Burz, D.S.; Breindel, L.; Shekhtman, A. The Inescapable Effects of Ribosomes on In-Cell NMR Spectroscopy and the Implications for Regulation of Biological Activity. Int. J. Mol. Sci. 2019, 20, 1297. https://doi.org/10.3390/ijms20061297
Burz DS, Breindel L, Shekhtman A. The Inescapable Effects of Ribosomes on In-Cell NMR Spectroscopy and the Implications for Regulation of Biological Activity. International Journal of Molecular Sciences. 2019; 20(6):1297. https://doi.org/10.3390/ijms20061297
Chicago/Turabian StyleBurz, David S., Leonard Breindel, and Alexander Shekhtman. 2019. "The Inescapable Effects of Ribosomes on In-Cell NMR Spectroscopy and the Implications for Regulation of Biological Activity" International Journal of Molecular Sciences 20, no. 6: 1297. https://doi.org/10.3390/ijms20061297