Articles | Volume 2, issue 1
https://doi.org/10.5194/mr-2-1-2021
© Author(s) 2021. This work is distributed under
the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Special issue:
https://doi.org/10.5194/mr-2-1-2021
© Author(s) 2021. This work is distributed under
the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Research article
| 
06 Jan 2021
Research article |
| 06 Jan 2021
Phosphoserine for the generation of lanthanide-binding sites on proteins for paramagnetic nuclear magnetic resonance spectroscopy
Sreelakshmi Mekkattu Tharayil
ARC Centre of Excellence for Innovations in Peptide and Protein
Science, Research School of Chemistry, Australian National University,
Canberra ACT 2601, Australia
Mithun Chamikara Mahawaththa
ARC Centre of Excellence for Innovations in Peptide and Protein
Science, Research School of Chemistry, Australian National University,
Canberra ACT 2601, Australia
Choy-Theng Loh
ARC Centre of Excellence for Innovations in Peptide and Protein
Science, Research School of Chemistry, Australian National University,
Canberra ACT 2601, Australia
present address: Hangzhou Wayland Bioscience Co. Ltd, Hangzhou
310030, PR China
Ibidolapo Adekoya
ARC Centre of Excellence for Innovations in Peptide and Protein
Science, Research School of Chemistry, Australian National University,
Canberra ACT 2601, Australia
Gottfried Otting
CORRESPONDING AUTHOR
ARC Centre of Excellence for Innovations in Peptide and Protein
Science, Research School of Chemistry, Australian National University,
Canberra ACT 2601, Australia
Related authors
Sreelakshmi Mekkattu Tharayil, Mithun C. Mahawaththa, Akiva Feintuch, Ansis Maleckis, Sven Ullrich, Richard Morewood, Michael J. Maxwell, Thomas Huber, Christoph Nitsche, Daniella Goldfarb, and Gottfried Otting
Magn. Reson., 3, 169–182, https://doi.org/10.5194/mr-3-169-2022, https://doi.org/10.5194/mr-3-169-2022, 2022
Short summary
Short summary
Having shown that tagging a protein at a single site with different lanthanoid complexes delivers outstanding structural information at a selected site of a protein (such as active sites and ligand binding sites), we now present a simple way by which different lanthanoid complexes can be assembled on a highly solvent-exposed cysteine residue. Furthermore, the chemical assembly is selective for selenocysteine, if a selenocysteine residue can be introduced into the protein of interest.
Damian Van Raad, Gottfried Otting, and Thomas Huber
Magn. Reson. Discuss., https://doi.org/10.5194/mr-2023-3, https://doi.org/10.5194/mr-2023-3, 2023
Preprint under review for MR
Short summary
Short summary
A novel cell-free protein synthesis system called eCells produces amino acids based on specific isotopes using low-cost precursors. The system selectively labels methyl groups; i.e valine and leucine with high efficiency. eCells achieve high levels of 13C incorporation and deuteration in protein preparations, making them suitable for NMR experiments of large protein complexes. It is easy to prepare and can be scaled up in volume, making it a promising tool for protein production and NMR studies.
Sreelakshmi Mekkattu Tharayil, Mithun C. Mahawaththa, Akiva Feintuch, Ansis Maleckis, Sven Ullrich, Richard Morewood, Michael J. Maxwell, Thomas Huber, Christoph Nitsche, Daniella Goldfarb, and Gottfried Otting
Magn. Reson., 3, 169–182, https://doi.org/10.5194/mr-3-169-2022, https://doi.org/10.5194/mr-3-169-2022, 2022
Short summary
Short summary
Having shown that tagging a protein at a single site with different lanthanoid complexes delivers outstanding structural information at a selected site of a protein (such as active sites and ligand binding sites), we now present a simple way by which different lanthanoid complexes can be assembled on a highly solvent-exposed cysteine residue. Furthermore, the chemical assembly is selective for selenocysteine, if a selenocysteine residue can be introduced into the protein of interest.
Henry W. Orton, Elwy H. Abdelkader, Lydia Topping, Stephen J. Butler, and Gottfried Otting
Magn. Reson., 3, 65–76, https://doi.org/10.5194/mr-3-65-2022, https://doi.org/10.5194/mr-3-65-2022, 2022
Short summary
Short summary
Installing a tag containing a paramagnetic metal ion on a protein can lead to large changes (pseudocontact shifts) in the resonances observed in NMR spectra. These are easily measured and contain valuable long-range structural information. The present work shows that a single tagging site furnished with different tags can be sufficient to localise atoms in proteins with high accuracy. In fact, this strategy works almost as well as the same number of tags distributed over multiple tagging sites.
Henry W. Orton, Iresha D. Herath, Ansis Maleckis, Shereen Jabar, Monika Szabo, Bim Graham, Colum Breen, Lydia Topping, Stephen J. Butler, and Gottfried Otting
Magn. Reson., 3, 1–13, https://doi.org/10.5194/mr-3-1-2022, https://doi.org/10.5194/mr-3-1-2022, 2022
Short summary
Short summary
This paper explores a method for determining the solution structure of a solvent-exposed polypeptide segment (the L3 loop), which is next to the active site of the penicillin-degrading enzyme IMP-1. Tagging three different sites on the protein with paramagnetic metal ions allowed positioning of the L3 loop with atomic resolution. It was found that the method was more robust when omitting data obtained with different metal ions if obtained with the same tag at the same tagging site.
Henry William Orton, Thomas Huber, and Gottfried Otting
Magn. Reson., 1, 1–12, https://doi.org/10.5194/mr-1-1-2020, https://doi.org/10.5194/mr-1-1-2020, 2020
Short summary
Short summary
Nuclear magnetic resonance is a technique that allows the measurement of the nanoscale distances between the atoms of a molecule. It is often relied upon for finding the structure of a molecule and how atoms are bonded, but measurements become inaccurate for large distances and therefore for large biological molecules. This research presents a new software for calculating the distances between nuclei and unpaired electrons which offers more accurate long-range distances in biological molecules.
Related subject area
Field: Liquid-state NMR | Topic: Applications – biological macromolecules
Facilitating the structural characterisation of non-canonical amino acids in biomolecular NMR
Site-selective generation of lanthanoid binding sites on proteins using 4-fluoro-2,6-dicyanopyridine
Imatinib disassembles the regulatory core of Abelson kinase by binding to its ATP site and not by binding to its myristoyl pocket
Localising nuclear spins by pseudocontact shifts from a single tagging site
Localising individual atoms of tryptophan side chains in the metallo-β-lactamase IMP-1 by pseudocontact shifts from paramagnetic lanthanoid tags at multiple sites
Fluorine NMR study of proline-rich sequences using fluoroprolines
Analysis of conformational exchange processes using methyl-TROSY-based Hahn echo measurements of quadruple-quantum relaxation
Anomalous amide proton chemical shifts as signatures of hydrogen bonding to aromatic sidechains
Rapid assessment of Watson–Crick to Hoogsteen exchange in unlabeled DNA duplexes using high-power SELOPE imino 1H CEST
High-affinity tamoxifen analogues retain extensive positional disorder when bound to calmodulin
Structural polymorphism and substrate promiscuity of a ribosome-associated molecular chaperone
Small-molecule inhibitors of the PDZ domain of Dishevelled proteins interrupt Wnt signalling
Real-time nuclear magnetic resonance spectroscopy in the study of biomolecular kinetics and dynamics
The long-standing relationship between paramagnetic NMR and iron–sulfur proteins: the mitoNEET example. An old method for new stories or the other way around?
Conformational features and ionization states of Lys side chains in a protein studied using the stereo-array isotope labeling (SAIL) method
Fragile protein folds: sequence and environmental factors affecting the equilibrium of two interconverting, stably folded protein conformations
Towards resolving the complex paramagnetic nuclear magnetic resonance (NMR) spectrum of small laccase: assignments of resonances to residue-specific nuclei
Sarah Kuschert, Martin Stroet, Yanni Ka-Yan Chin, Anne Claire Conibear, Xinying Jia, Thomas Lee, Christian Reinhard Otto Bartling, Kristian Strømgaard, Peter Güntert, Karl Johan Rosengren, Alan Edward Mark, and Mehdi Mobli
Magn. Reson., 4, 57–72, https://doi.org/10.5194/mr-4-57-2023, https://doi.org/10.5194/mr-4-57-2023, 2023
Short summary
Short summary
The 20 genetically encoded amino acids provide the basis for most proteins and peptides that make up the machinery of life. This limited repertoire is vastly expanded by the introduction of non-canonical amino acids (ncAAs). Studying the structure of protein-containing ncAAs requires new computational representations that are compatible with existing modelling software. We have developed an online tool for this to aid future structural studies of this class of complex biopolymer.
Sreelakshmi Mekkattu Tharayil, Mithun C. Mahawaththa, Akiva Feintuch, Ansis Maleckis, Sven Ullrich, Richard Morewood, Michael J. Maxwell, Thomas Huber, Christoph Nitsche, Daniella Goldfarb, and Gottfried Otting
Magn. Reson., 3, 169–182, https://doi.org/10.5194/mr-3-169-2022, https://doi.org/10.5194/mr-3-169-2022, 2022
Short summary
Short summary
Having shown that tagging a protein at a single site with different lanthanoid complexes delivers outstanding structural information at a selected site of a protein (such as active sites and ligand binding sites), we now present a simple way by which different lanthanoid complexes can be assembled on a highly solvent-exposed cysteine residue. Furthermore, the chemical assembly is selective for selenocysteine, if a selenocysteine residue can be introduced into the protein of interest.
Stephan Grzesiek, Johannes Paladini, Judith Habazettl, and Rajesh Sonti
Magn. Reson., 3, 91–99, https://doi.org/10.5194/mr-3-91-2022, https://doi.org/10.5194/mr-3-91-2022, 2022
Short summary
Short summary
We show here that binding of the anticancer drug imatinib to the ATP site of Abelson kinase and not binding to its allosteric site coincides with the opening of the kinase regulatory core at nanomolar concentrations. This has implications for the understanding of Abelson’s kinase regulation and activity during medication as well as for the design of new Abelson kinase inhibitors.
Henry W. Orton, Elwy H. Abdelkader, Lydia Topping, Stephen J. Butler, and Gottfried Otting
Magn. Reson., 3, 65–76, https://doi.org/10.5194/mr-3-65-2022, https://doi.org/10.5194/mr-3-65-2022, 2022
Short summary
Short summary
Installing a tag containing a paramagnetic metal ion on a protein can lead to large changes (pseudocontact shifts) in the resonances observed in NMR spectra. These are easily measured and contain valuable long-range structural information. The present work shows that a single tagging site furnished with different tags can be sufficient to localise atoms in proteins with high accuracy. In fact, this strategy works almost as well as the same number of tags distributed over multiple tagging sites.
Henry W. Orton, Iresha D. Herath, Ansis Maleckis, Shereen Jabar, Monika Szabo, Bim Graham, Colum Breen, Lydia Topping, Stephen J. Butler, and Gottfried Otting
Magn. Reson., 3, 1–13, https://doi.org/10.5194/mr-3-1-2022, https://doi.org/10.5194/mr-3-1-2022, 2022
Short summary
Short summary
This paper explores a method for determining the solution structure of a solvent-exposed polypeptide segment (the L3 loop), which is next to the active site of the penicillin-degrading enzyme IMP-1. Tagging three different sites on the protein with paramagnetic metal ions allowed positioning of the L3 loop with atomic resolution. It was found that the method was more robust when omitting data obtained with different metal ions if obtained with the same tag at the same tagging site.
Davy Sinnaeve, Abir Ben Bouzayene, Emile Ottoy, Gert-Jan Hofman, Eva Erdmann, Bruno Linclau, Ilya Kuprov, José C. Martins, Vladimir Torbeev, and Bruno Kieffer
Magn. Reson., 2, 795–813, https://doi.org/10.5194/mr-2-795-2021, https://doi.org/10.5194/mr-2-795-2021, 2021
Short summary
Short summary
Fluorine NMR was used to study the interaction between a proline-rich peptide and a SH3 domain using 4S- and 4R-fluorinated prolines whose potential as NMR probes has not been exploited yet. We present a comprehensive study addressing several aspects to be considered when using these residues as NMR probes, including relaxation and dynamics. We show that their conformational bias may be used to modulate the kinetics of protein binding to proline-rich motifs.
Christopher A. Waudby and John Christodoulou
Magn. Reson., 2, 777–793, https://doi.org/10.5194/mr-2-777-2021, https://doi.org/10.5194/mr-2-777-2021, 2021
Short summary
Short summary
We describe a suite of experiments that exploit field-dependent relaxation measurements of four-spin transitions in methyl groups to characterise chemical exchange processes and which can be used as an alternative or complement to CPMG relaxation dispersion measurements. We show that these four-spin transitions benefit from the methyl TROSY effect and so provide a unique combination of slow intrinsic relaxation and high sensitivity to chemical exchange.
Kumaran Baskaran, Colin W. Wilburn, Jonathan R. Wedell, Leonardus M. I. Koharudin, Eldon L. Ulrich, Adam D. Schuyler, Hamid R. Eghbalnia, Angela M. Gronenborn, and Jeffrey C. Hoch
Magn. Reson., 2, 765–775, https://doi.org/10.5194/mr-2-765-2021, https://doi.org/10.5194/mr-2-765-2021, 2021
Short summary
Short summary
The Biological Magnetic Resonance Data Bank (BMRB) has been used to identify overall trends, for example, the relationship between chemical shift and backbone conformation. The BMRB archive has grown so that statistical outliers are sufficiently numerous to afford insights into unusual or unique structural features in proteins. We analyze amide proton chemical shift outliers to gain insights into the occurrence of hydrogen bonds between an amide NH and the p-pi cloud of aromatic sidechains.
Bei Liu, Atul Rangadurai, Honglue Shi, and Hashim M. Al-Hashimi
Magn. Reson., 2, 715–731, https://doi.org/10.5194/mr-2-715-2021, https://doi.org/10.5194/mr-2-715-2021, 2021
Short summary
Short summary
There is growing interest in mapping exchange dynamics between Watson–Crick and Hoogsteen conformations across different DNA contexts. However, current methods are ill-suited for measurements at a large scale because they require isotopically enriched samples. We report that Hoogsteen dynamics can be measured on unlabeled samples using 1H CEST experiments, which have higher throughput and lower cost relative to conventional methods and also provide new insights into Hoogsteen dynamics.
Lilia Milanesi, Clare R. Trevitt, Brian Whitehead, Andrea M. Hounslow, Salvador Tomas, Laszlo L. P. Hosszu, Christopher A. Hunter, and Jonathan P. Waltho
Magn. Reson., 2, 629–642, https://doi.org/10.5194/mr-2-629-2021, https://doi.org/10.5194/mr-2-629-2021, 2021
Short summary
Short summary
The overall aim of the study is to provide a basis from which to improve the ability of tamoxifen family drugs to reduce the activity of a secondary target protein, calmodulin, during tumour development. The main conclusion is that the binding of a tamoxifen analogue is quite unlike that of other anti-calmodulin compounds in that two drug molecules bring the two domains of calmodulin into close proximity, but they are not fixed in orientation relative to the protein.
Chih-Ting Huang, Yei-Chen Lai, Szu-Yun Chen, Meng-Ru Ho, Yun-Wei Chiang, and Shang-Te Danny Hsu
Magn. Reson., 2, 375–386, https://doi.org/10.5194/mr-2-375-2021, https://doi.org/10.5194/mr-2-375-2021, 2021
Short summary
Short summary
Trigger factor (TF) is a conserved bacterial molecular chaperone that exists in a monomer–dimer equilibrium in solution. It binds to the ribosome as a monomer to facilitate folding of nascent polypeptide chains. We showed that dimeric TF exhibits distinct domain dynamics and conformational polymorphism and that TF contains multiple substrate binding sites that are only accessible in its monomeric form. The equilibrium of TF in different oligomeric states may serve as a regulatory mechanism.
Nestor Kamdem, Yvette Roske, Dmytro Kovalskyy, Maxim O. Platonov, Oleksii Balinskyi, Annika Kreuchwig, Jörn Saupe, Liang Fang, Anne Diehl, Peter Schmieder, Gerd Krause, Jörg Rademann, Udo Heinemann, Walter Birchmeier, and Hartmut Oschkinat
Magn. Reson., 2, 355–374, https://doi.org/10.5194/mr-2-355-2021, https://doi.org/10.5194/mr-2-355-2021, 2021
Short summary
Short summary
The Wnt signalling pathway plays a major role in prevention of cancer, whereby the protein Dishevelled connects from the transmembrane receptor Frizzled to downstream effectors via its PDZ domain. Here, cycles of chemical synthesis and structural biology are applied to develop PDZ ligands that block the Frizzled–Dishevelled interaction using NMR for screening, in ligand development, and for deriving structure–activity relationships. Cellular reporter assays demonstrate their efficacy.
György Pintér, Katharina F. Hohmann, J. Tassilo Grün, Julia Wirmer-Bartoschek, Clemens Glaubitz, Boris Fürtig, and Harald Schwalbe
Magn. Reson., 2, 291–320, https://doi.org/10.5194/mr-2-291-2021, https://doi.org/10.5194/mr-2-291-2021, 2021
Short summary
Short summary
The folding, refolding and misfolding of biomacromolecules including proteins, DNA and RNA is an important area of biophysical research to understand functional and disease states of a cell. NMR spectroscopy provides detailed insight, with both high time and atomic resolution. These experiments put stringent requirements on signal-to-noise for often irreversible folding reactions. The review describes methodological approaches and highlights key applications.
Francesca Camponeschi, Angelo Gallo, Mario Piccioli, and Lucia Banci
Magn. Reson., 2, 203–221, https://doi.org/10.5194/mr-2-203-2021, https://doi.org/10.5194/mr-2-203-2021, 2021
Short summary
Short summary
The iron–sulfur cluster binding properties of human mitoNEET have been investigated by 1D and 2D 1H paramagnetic NMR spectroscopy. The NMR spectra of both oxidized and reduced mitoNEET are significantly different from those reported previously for other [Fe2S2] proteins. Our findings revealed the unique electronic properties of mitoNEET and suggests that the specific electronic structure of the cluster possibly drives the functional properties of different [Fe2S2] proteins.
Mitsuhiro Takeda, Yohei Miyanoiri, Tsutomu Terauchi, and Masatsune Kainosho
Magn. Reson., 2, 223–237, https://doi.org/10.5194/mr-2-223-2021, https://doi.org/10.5194/mr-2-223-2021, 2021
Short summary
Short summary
Although both the hydrophobic aliphatic chain and hydrophilic ζ-amino group of the lysine side chain presumably contribute to the structures and functions of proteins, the dual nature of the lysine residue has not been fully understood yet, due to the lack of appropriate methods to acquire comprehensive information on its long consecutive methylene chain at the atomic scale. We describe herein a novel strategy to address the current situation using nuclear magnetic resonance spectroscopy.
Xingjian Xu, Igor Dikiy, Matthew R. Evans, Leandro P. Marcelino, and Kevin H. Gardner
Magn. Reson., 2, 63–76, https://doi.org/10.5194/mr-2-63-2021, https://doi.org/10.5194/mr-2-63-2021, 2021
Short summary
Short summary
While most proteins adopt one conformation, several interconvert between two or more very different structures. Knowing how sequence changes and small-molecule binding can control this behavior is essential for both understanding biology and inspiring new “molecular switches” which can control cellular pathways. This work contributes by examining these topics in the ARNT protein, showing that features of both the folded and unfolded states contribute to the interconversion process.
Rubin Dasgupta, Karthick B. S. S. Gupta, Huub J. M. de Groot, and Marcellus Ubbink
Magn. Reson., 2, 15–23, https://doi.org/10.5194/mr-2-15-2021, https://doi.org/10.5194/mr-2-15-2021, 2021
Short summary
Short summary
A method is demonstrated that can help in sequence-specific NMR signal assignment to nuclear spins near a strongly paramagnetic metal in an enzyme. A combination of paramagnetically tailored NMR experiments and second-shell mutagenesis was used to attribute previously observed chemical exchange processes in the active site of laccase to specific histidine ligands. The signals of nuclei close to the metal can be used as spies to unravel the role of motions in the catalytic process.
Cited articles
Abdelkader, E. H., Yao, X., Feintuch, A., Adams, L. A., Aurelio, L., Graham,
B., Goldfarb, D., and Otting, G.: Pulse EPR-enabled interpretation of scarce
pseudocontact shifts induced by lanthanide binding tags, J. Biomol. NMR, 64,
39–51, https://doi.org/10.1007/s10858-015-0003-z, 2016.
Baneyx, F. and Mujacic, M.: Recombinant protein folding and misfolding in
Escherichia coli, Nat. Biotechnol., 22, 1399–1408, https://doi.org/10.1038/nbt1029, 2004.
Barthelmes, D., Gränz, M., Barthelmes, K., Allen, K. N., Imperiali, B.,
Prisner, T., and Schwalbe, H.: Encoded loop-lanthanide-binding tags for
long-range distance measurements in proteins by NMR and EPR spectroscopy, J.
Biomol. NMR, 63, 275–282, https://doi.org/10.1007/s10858-015-9984-x, 2015.
Barthelmes, D., Barthelmes, K., Schnorr, K., Jonker, H. R. A., Bodmer, B.,
Allen, K. N., Imperiali, B., and Schwalbe, H.: Conformational dynamics and
alignment properties of loop lanthanide-binding-tags (LBTs) studied in
interleukin-1β, J. Biomol. NMR, 68, 187–194,
https://doi.org/10.1007/s10858-017-0118-5, 2017.
Barthelmes, K., Reynolds, A. M., Peisach, E., Jonker, H. R. A., DeNunzio, N.
J., Allen, K. N., Imperiali, B., and Schwalbe, H.: Engineering encodable
lanthanide-binding tags into loop regions of proteins, J. Am. Chem. Soc.,
133, 808–819, https://doi.org/10.1021/ja104983t, 2011.
Bertini, I., Janik, M. B. L., Lee, Y.-M., Luchinat, C., and Rosato, A.:
Magnetic susceptibility tensor anisotropies for a lanthanide ion series in a
fixed protein matrix, J. Am. Chem. Soc., 123, 4181–4188,
https://doi.org/10.1021/ja0028626, 2001.
Bleaney, B.: Nuclear magnetic resonance shifts in solution due to lanthanide
ions, J. Magn. Reson., 8, 91–100, https://doi.org/10.1016/0022-2364(72)90027-3,
1972.
Bryson, D. I., Fan, C., Guo, L.-T., Miller, C., Söll, D., and Liu, D.
R.: Continuous directed evolution of aminoacyl-tRNA synthetases, Nat. Chem.
Biol., 13, 1253–1260, https://doi.org/10.1038/nchembio.2474, 2017.
Dumas, A., Lercher, L., Spicer, C. D., and Davis, B. G.: Designing logical
codon reassignment – Expanding the chemistry in biology, Chem. Sci., 6,
50–69, https://doi.org/10.1039/C4SC01534G, 2014.
Fenwick, R. B., Esteban-Martín, S., Richter, B., Lee, D., Walter, K. F.
A., Milovanovic, D., Becker, S., Lakomek, N. A., Griesinger, C., and
Salvatella, X.: Weak long-range correlated motions in a surface patch of
ubiquitin involved in molecular recognition, J. Am. Chem. Soc., 133,
10336–10339, https://doi.org/10.1021/ja200461n, 2011.
Gallagher, T., Alexander, P., Bryan, P., and Gilliland, G. L.: Two crystal
structures of the B1 immunoglobulin-binding domain of streptococcal protein
G and comparison with NMR, Biochemistry, 33, 4721–4729,
https://doi.org/10.1021/bi00181a032, 1994.
Hass, M. A. S., Keizers, P. H. J., Blok, A., Hiruma, Y., and Ubbink, M.:
Validation of a lanthanide tag for the analysis of protein dynamics by
paramagnetic NMR spectroscopy, J. Am. Chem. Soc., 132, 9952–9953,
https://doi.org/10.1021/ja909508r, 2010.
Jia, X., Yagi, H., Su, X.-C., Stanton-Cook, M., Huber, T., and Otting, G.:
Engineering [Ln(DPA)3]3− binding sites in proteins: a widely
applicable method for tagging proteins with lanthanide ions, J. Biomol. NMR,
50, 411–420, https://doi.org/10.1007/s10858-011-9529-x, 2011.
Jones, D. H., Cellitti, S. E., Hao, X., Zhang, Q., Jahnz, M., Summerer, D.,
Schultz, P. G., Uno, T., and Geierstanger, B. H.: Site-specific labeling of
proteins with NMR-active unnatural amino acids, J. Biomol. NMR, 46, 89–100,
https://doi.org/10.1007/s10858-009-9365-4, 2009.
Joss, D. and Häussinger, D.: Design and applications of lanthanide
chelating tags for pseudocontact shift NMR spectroscopy with
biomacromolecules, Prog. Nucl. Mag. Res. Sp., 114–115, 284–312,
https://doi.org/10.1016/j.pnmrs.2019.08.002, 2019.
Keizers, P. H. J. and Ubbink, M.: Paramagnetic tagging for protein structure
and dynamics analysis, Prog. Nucl. Mag. Res. Sp., 58, 88–96,
https://doi.org/10.1016/j.pnmrs.2010.08.001, 2011.
Keller, S., Vargas, C., Zhao, H., Piszczek, G., Brautigam, C. A., and
Schuck, P.: High-precision isothermal titration calorimetry with automated
peak-shape analysis, Anal. Chem., 84, 5066–5073, https://doi.org/10.1021/ac3007522,
2012.
Lammers, C., Hahn, L. E., and Neumann, H.: Optimized plasmid systems for the
incorporation of multiple different unnatural amino acids by evolved
orthogonal ribosomes, ChemBioChem, 15, 1800–1804,
https://doi.org/10.1002/cbic.201402033, 2014.
Lee, S., Oh, S., Yang, A., Kim, J., Söll, D., Lee, D., and Park, H.-S.:
A facile strategy for selective incorporation of phosphoserine into
histones, Angew. Chem. Int. Edit., 52, 5771–5775,
https://doi.org/10.1002/anie.201300531, 2013.
Loh, C. T., Ozawa, K., Tuck, K. L., Barlow, N., Huber, T., Otting, G., and
Graham, B.: Lanthanide tags for site-specific ligation to an unnatural amino
acid and generation of pseudocontact shifts in proteins, Bioconjugate Chem.,
24, 260–268, https://doi.org/10.1021/bc300631z, 2013.
Loh, C.-T., Graham, B., Abdelkader, E. H., Tuck, K. L., and Otting, G.:
Generation of pseudocontact shifts in proteins with lanthanides using small
“clickable” nitrilotriacetic acid and iminodiacetic acid tags, Chem. Eur.
J., 21, 5084–5092, https://doi.org/10.1002/chem.201406274, 2015.
Mekkattu Tharayil, S., Mahawaththa, M. C., Adekoya, I., Otting, G.: NMR spectra, Australian National University, https://doi.org/10.25911/5fc5bd5f0f872, 2020.
Morgan, K.: Plasmids 101: origin of replication, available at:
https://blog.addgene.org/plasmid-101-origin-of-replication (last access: 27 October 2020), 2014.
Neylon, C., Brown, S. E., Kralicek, A. V., Miles, C. S., Love, C. A., and
Dixon, N. E.: Interaction of the Escherichia coli replication terminator protein (Tus) with DNA: A model derived from DNA-Binding studies of mutant proteins by surface plasmon resonance, Biochemistry, 39, 11989–11999, https://doi.org/10.1021/bi001174w, 2000.
Nguyen, T. H. D., Ozawa, K., Stanton-Cook, M., Barrow, R., Huber, T., and
Otting, G.: Generation of pseudocontact shifts in protein NMR spectra with a
genetically encoded cobalt(II)-binding amino acid, Angew. Chem. Int. Edit.,
50, 692–694, https://doi.org/10.1002/anie.201005672, 2011.
Nitsche, C. and Otting, G.: Pseudocontact shifts in biomolecular NMR using
paramagnetic metal tags, Prog. Nucl. Mag. Res. Sp., 98–99, 20–49, https://doi.org/10.1016/j.pnmrs.2016.11.001, 2017.
Orton, H. W., Huber, T., and Otting, G.: Paramagpy: software for fitting
magnetic susceptibility tensors using paramagnetic effects measured in NMR
spectra, Magn. Reson., 1, 1–12, doi:https://doi.org/10.5194/mr-1-1-2020,
2020.
Otting, G.: Prospects for lanthanides in structural biology by NMR, J.
Biomol. NMR, 42, 1–9, https://doi.org/10.1007/s10858-008-9256-0, 2008.
Parigi, G. and Luchinat, C.: NMR consequences of the nucleus-electron spin
interactions, in: Paramagnetism in Experimental Biomolecular NMR, edited by:
Luchinat, C., Parigi, G., and Ravera, E., RSC Publishing, Cambridge, United
Kingdom, 1–41, https://doi.org/10.1039/9781788013291-00001, 2018.
Park, H.-S., Hohn, M. J., Umehara, T., Guo, L.-T., Osborne, E. M., Benner,
J., Noren, C. J., Rinehart, J., and Söll, D.: Expanding the genetic code
of Escherichia coli with phosphoserine, Science, 333, 1151–1154,
https://doi.org/10.1126/science.1207203, 2011.
Pirman, N. L., Barber, K. W., Aerni, H. R., Ma, N. J., Haimovich, A. D.,
Rogulina, S., Isaacs, F. J., and Rinehart, J.: A flexible codon in
genomically recoded Escherichia coli permits programmable protein phosphorylation, Nat. Commun., 6, 8130, https://doi.org/10.1038/ncomms9130, 2015.
Pott, M., Schmidt, M. J., and Summerer, D.: Evolved sequence contexts for
highly efficient amber suppression with noncanonical amino acids, ACS Chem.
Biol., 9, 2815–2822, https://doi.org/10.1021/cb5006273, 2014.
Qi, R. and Otting, G.: Mutant T4 DNA polymerase for easy cloning and
mutagenesis, PLOS One, 14, e0211065, https://doi.org/10.1371/journal.pone.0211065, 2019.
Saio, T. and Ishimori, K.: Accelerating structural life science by
paramagnetic lanthanide probe methods, BBA-Gen. Subjects,
1864, 129332, https://doi.org/10.1016/j.bbagen.2019.03.018, 2020.
Saio, T., Ogura, K., Yokochi, M., Kobashigawa, Y., and Inagaki, F.:
Two-point anchoring of a lanthanide-binding peptide to a target protein
enhances the paramagnetic anisotropic effect, J. Biomol. NMR, 44, 157–166,
https://doi.org/10.1007/s10858-009-9325-z, 2009.
Saio, T., Yokochi, M., Kumeta, H., and Inagaki, F.: PCS-based structure
determination of protein–protein complexes, J. Biomol. NMR, 46, 271–280,
https://doi.org/10.1007/s10858-010-9401-4, 2010.
Saio, T., Ogura, K., Shimizu, K., Yokochi, M., Burke, T. R., and Inagaki,
F.: An NMR strategy for fragment-based ligand screening utilizing a
paramagnetic lanthanide probe, J. Biomol. NMR, 51, 395–408,
https://doi.org/10.1007/s10858-011-9566-5, 2011.
Shishmarev, D. and Otting, G.: How reliable are pseudocontact shifts induced
in proteins and ligands by mobile paramagnetic metal tags? A modelling
study, J. Biomol. NMR, 56, 203–216, https://doi.org/10.1007/s10858-013-9738-6, 2013.
Su, X.-C. and Chen, J.-L.: Site-specific tagging of proteins with paramagnetic ions for determination of protein structures in solution and in cells, Acc. Chem. Res., 52, 1675–1686, https://doi.org/10.1021/acs.accounts.9b00132, 2019.
Su, X.-C. and Otting, G.: Paramagnetic labelling of proteins and
oligonucleotides for NMR, J. Biomol. NMR, 46, 101–112,
https://doi.org/10.1007/s10858-009-9331-1, 2010.
Swarbrick, J. D., Ung, P., Su, X.-C., Maleckis, A., Chhabra, S., Huber, T.,
Otting, G., and Graham, B.: Engineering of a bis-chelator motif into a
protein α-helix for rigid lanthanide binding and paramagnetic NMR
spectroscopy, Chem. Commun., 47, 7368–7370, https://doi.org/10.1039/C1CC11893E, 2011.
Swarbrick, J. D., Ung, P., Dennis, M. L., Lee, M. D., Chhabra, S., and
Graham, B.: Installation of a rigid EDTA-like motif into a protein α-helix for paramagnetic NMR spectroscopy with cobalt(II) ions, Chem. Eur.
J., 22, 1228–1232, https://doi.org/10.1002/chem.201503139, 2016.
Welegedara, A. P., Yang, Y., Lee, M. D., Swarbrick, J. D., Huber, T.,
Graham, B., Goldfarb, D., and Otting, G.: Double-arm lanthanide tags deliver
narrow Gd3+–Gd3+ distance distributions in double
electron–electron resonance (DEER) measurements, Chem.-Eur. J., 23,
11694–11702, https://doi.org/10.1002/chem.201702521, 2017.
Xie, Y., Jiang, Y., and Ben-Amotz, D.: Detection of amino acid and peptide
phosphate protonation using Raman spectroscopy, Anal. Biochem., 343,
223–230, https://doi.org/10.1016/j.ab.2005.05.038, 2005.
Yagi, H., Loscha, K. V., Su, X.-C., Stanton-Cook, M., Huber, T., and Otting,
G.: Tunable paramagnetic relaxation enhancements by [Gd(DPA)3]3−
for protein structure analysis, J. Biomol. NMR, 47, 143–153,
https://doi.org/10.1007/s10858-010-9416-x, 2010.
Yang, A., Ha, S., Ahn, J., Kim, R., Kim, S., Lee, Y., Kim, J., Söll, D.,
Lee, H.-Y., and Park, H.-S.: A chemical biology route to site-specific
authentic protein modifications, Science, 354, 623–626,
https://doi.org/10.1126/science.aah4428, 2016.
Short summary
A new way is presented for creating lanthanide binding sites on proteins using site-specifically introduced phosphoserine residues. The paramagnetic effects of lanthanides generate long-range effects, which contain structural information and are readily measured by NMR spectroscopy. Excellent correlations between experimentally observed and back-calculated pseudocontact shifts attest to very good immobilization of the lanthanide ions relative to the proteins.
A new way is presented for creating lanthanide binding sites on proteins using site-specifically...
Special issue