Articles | Volume 6, issue 2
https://doi.org/10.5194/mr-6-157-2025
© Author(s) 2025. This work is distributed under
the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
https://doi.org/10.5194/mr-6-157-2025
© Author(s) 2025. This work is distributed under
the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Research article
| 
16 Jul 2025
Research article |
| 16 Jul 2025
| 16 Jul 2025
3D-printed microcell for protein NMR at high ionic strengths and small sample volumes
Tayeb Kakeshpour
CORRESPONDING AUTHOR
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, USA
Martin D. Gelenter
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, USA
Jinfa Ying
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, USA
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, USA
Related authors
No articles found.
Yulia Pustovalova, Frank Delaglio, D. Levi Craft, Haribabu Arthanari, Ad Bax, Martin Billeter, Mark J. Bostock, Hesam Dashti, D. Flemming Hansen, Sven G. Hyberts, Bruce A. Johnson, Krzysztof Kazimierczuk, Hengfa Lu, Mark Maciejewski, Tomas M. Miljenović, Mehdi Mobli, Daniel Nietlispach, Vladislav Orekhov, Robert Powers, Xiaobo Qu, Scott Anthony Robson, David Rovnyak, Gerhard Wagner, Jinfa Ying, Matthew Zambrello, Jeffrey C. Hoch, David L. Donoho, and Adam D. Schuyler
Magn. Reson., 2, 843–861, https://doi.org/10.5194/mr-2-843-2021, https://doi.org/10.5194/mr-2-843-2021, 2021
Short summary
Short summary
We present the ongoing work of a large, community initiative to establish standards for the processing of nonuniformly sampled NMR experiments. The NUScon software, contest, and archive of spectral evaluation data provide a comprehensive platform for addressing the most challenging questions related to NUS experiments. We will run annual contests and generate a database of results, which will empower us in guiding the NUS community towards a set of best practices.
Angus J. Robertson, Jinfa Ying, and Ad Bax
Magn. Reson., 2, 129–138, https://doi.org/10.5194/mr-2-129-2021, https://doi.org/10.5194/mr-2-129-2021, 2021
Short summary
Short summary
NMR study of large proteins such as SARS-CoV-2 Main Protease can be challenging when exchange broadening, multiple stable conformations, and back-exchanging the fully deuterated chain pose problems. We demonstrate that 4D NMR, including an extension of 3D NOE-NOE spectroscopy, provides an effective tool for spectral analysis. In combination with X-ray coordinates, the 4D NMR data are particularly useful for extending and validating assignments and for probing structural features.
Related subject area
Field: Liquid-state NMR | Topic: Applications – biological macromolecules
NMR side-chain assignments of the Crimean–Congo hemorrhagic fever virus glycoprotein n cytosolic domain
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
Phosphoserine for the generation of lanthanide-binding sites on proteins for paramagnetic nuclear magnetic resonance spectroscopy
Louis Brigandat, Maëlys Laux, Caroline Marteau, Laura Cole, Anja Böckmann, Lauriane Lecoq, Marie-Laure Fogeron, and Morgane Callon
Magn. Reson., 5, 95–101, https://doi.org/10.5194/mr-5-95-2024, https://doi.org/10.5194/mr-5-95-2024, 2024
Short summary
Short summary
We used NMR to sequentially assign the side-chain resonances of the cytosolic domain of glycoprotein n of the Crimean–Congo hemorrhagic fever virus. The combination of cell-free protein synthesis with high-field NMR and artificial intelligence approaches facilitated a time- and effort-efficient approach. Our results will be harnessed to study the membrane-bound form of the domain and its interactions with virulence factors, which will ultimately help to understand their role in disease.
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.
Sreelakshmi Mekkattu Tharayil, Mithun Chamikara Mahawaththa, Choy-Theng Loh, Ibidolapo Adekoya, and Gottfried Otting
Magn. Reson., 2, 1–13, https://doi.org/10.5194/mr-2-1-2021, https://doi.org/10.5194/mr-2-1-2021, 2021
Short summary
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.
Cited articles
Barbara, T. M.: Cylindrical Demagnetization Fields and Microprobe Design in High-Resolution NMR, J. Magn. Reson. Ser. A, 109, 265–269, https://doi.org/10.1006/jmra.1994.1168, 1994.
Barbara, T. M.: NMR Probes for Small Sample Volumes, Encyclopedia of Magnetic Resonance, edited by: Harris, R. K. and Wasylishen, R., John Wiley, Chichester, https://doi.org/10.1002/9780470034590.emrstm1084, 2009.
Bartels, T., Choi, J. G., and Selkoe, D. J.: Alpha-Synuclein Occurs Physiologically as a Helically Folded Tetramer that Resists Aggregation, Nature, 477, 107–110, https://doi.org/10.1038/nature10324, 2011.
Bretthorst, G. L.: Automatic Phasing of MR Images. Part II: Voxel-wise Phase Estimation, J. Magn. Reson., 191, 193–201, https://doi.org/10.1016/j.jmr.2007.12.011, 2008.
Delaglio, F., Grzesiek, S., Vuister, G. W., Zhu, G., Pfeifer, J., and Bax, A.: NMRPipe: a Multidimensional Spectral Processing System Based on UNIX Pipes, J. Biomol. NMR, 6, 277–293, https://doi.org/10.1007/BF00197809, 1995.
Evans, D. F.: The Determination of the Paramagnetic Susceptibility of Substances in Solution by Nuclear Magnetic Resonance, J. Chem. Soc., 1959, 2003–2005, https://doi.org/10.1039/jr9590002003, 1959.
Freeman, R., Kempsell, S. P., and Levitt, M. H.: Radiofrequency Pulse Sequences which Compensate their Own Imperfections, J. Magn. Reson., 38, 453–479, https://doi.org/10.1016/0022-2364(80)90327-3, 1980.
Geen, H. and Freeman, R.: Band-selective Radiofrequency Pulses, J. Magn. Reson., 93, 93–141, https://doi.org/10.1016/0022-2364(91)90034-Q, 1991.
Gelenter, M. and Bax, A.: Recombinant Expression and Chemical Amidation of Isotopically Labeled Native Melittin, J. Am. Chem. Soc., 145, 3850–3854, https://doi.org/10.1021/jacs.2c12631, 2023.
Gelenter, M. D., Yau, W. M., Anfinrud, P. A., and Bax, A.: From Milliseconds to Minutes: Melittin Self-Assembly from Concerted Non-Equilibrium Pressure-Jump and Equilibrium Relaxation Nuclear Magnetic Resonance, J. Phys. Chem. Lett., 15, 1930–1935, https://doi.org/10.1021/acs.jpclett.3c03563, 2024.
Gutiérrez-Mejía, F. and Ruiz-Suárez, J. C.: AC Magnetic Susceptibility at Medium Frequencies Suggests a Paramagnetic Behavior of Pure Water, J. Magn. Magn. Mater., 324, 1129–1132, https://doi.org/10.1016/j.jmmm.2011.10.035, 2012.
Hale, W., Rossetto, G., Greenhalgh, R., Finch, G., and Utz, M.: High-resolution Nuclear Magnetic Resonance Spectroscopy in Microfluidic Droplets, Lab on a Chip, 18, 3018–3024, https://doi.org/10.1039/c8lc00712h, 2018.
Hizawa, T., Takahashi, M., and Iwase, E.: Sample Shape Design for a Micro-volume NMR Spectroscopy, Micro Nano Lett., 12, 550–553, https://doi.org/10.1049/mnl.2017.0138, 2017.
Hoffman, R.: Magnetic Susceptibility Measurement by NMR: 2. The Magnetic Susceptibility of NMR Solvents and their Chemical Shifts, J. Magn. Reson., 335, 107105, https://doi.org/10.1016/j.jmr.2021.107105, 2022.
Hoffman, R. E.: Magnetic Susceptibility Measurement by NMR: 1. The Temperature Dependence of TMS, J. Magn. Reson., 312, 106689, https://doi.org/10.1016/j.jmr.2020.106689, 2020.
Johnson, M., Coulton, A. T., Geeves, M. A., and Mulvihill, D. P.: Targeted Amino-Terminal Acetylation of Recombinant Proteins in E. coli, PLoS One, 5, e15801, https://doi.org/10.1371/journal.pone.0015801, 2010.
Kakeshpour, T., Gelenter, M., Ying, J., and Bax, A.: 3D-printed microcell for protein NMR at high ionic strengths and small sample volumes (STL file), Zenodo [code], https://doi.org/10.5281/zenodo.15064675, 2025a.
Kakeshpour, T., Gelenter, M., Ying, J., and Bax, A.: 3D-printed microcell for protein NMR at high ionic strengths and small sample volumes (Python scripts), Zenodo [data set], https://doi.org/10.5281/zenodo.15858288, 2025b.
Kang, L., Moriarty, G. M., Woods, L. A., Ashcroft, A. E., Radford, S. E., and Baum, J.: N-Terminal Acetylation of Alpha-Synuclein Induces Increased Transient Helical Propensity and Decreased Aggregation Rates in the Intrinsically Disordered Monomer, Protein Sci., 21, 911–917, https://doi.org/10.1002/pro.2088, 2012.
Kay, L. E., Keifer, P., and Saarinen, T.: Pure Absorption Gradient-Enhanced Heteronuclear Single Quantum Correlation Spectroscopy with Improved Sensitivity, J. Am. Chem. Soc., 114, 10663–10665, https://doi.org/10.1021/ja00052a088, 1992.
Long, Z., Ruthford, J., and Opella, S. J.: 3D Printed Sample Tubes for Solid-State NMR Experiments, J. Magn. Reson., 327, 106957, https://doi.org/10.1016/j.jmr.2021.106957, 2021.
Maltsev, A. S., Ying, J. F., and Bax, A.: Impact of N-Terminal Acetylation of α-Synuclein on Its Random Coil and Lipid Binding Properties, Biochemistry, 51, 5004–5013, https://doi.org/10.1021/bi300642h, 2012.
Mangas-Florencio, L., Herrero-Gómez, A., Eills, J., Azagra, M., Batlló-Rius, M., and Marco-Rius, I.: A DIY Bioreactor for in Situ Metabolic Tracking in 3D Cell Models via Hyperpolarized 13C NMR Spectroscopy, Anal. Chem., 97, 1594–1602, https://doi.org/10.1021/acs.analchem.4c04183, 2025.
Manu, V. S., Olivieri, C., and Veglia, G.: AI-designed NMR Spectroscopy RF pulses for Fast Acquisition at High and Ultra-High Magnetic Fields, Nat. Commun., 14, 4144, https://doi.org/10.1038/s41467-023-39581-4, 2023.
Palmer, A. G., Cavanagh, J., Wright, P. E., and Rance, M.: Sensitivity Improvement in Proton-Detected 2-Dimensional Heteronuclear Correlation Nmr-Spectroscopy, J. Magn. Reson., 93, 151–170, https://doi.org/10.1016/0022-2364(91)90036-S, 1991.
Pereira, D., Sardo, M., Marín-Montesinos, I., and Mafra, L.: One-Shot Resin 3D-Printed Stators for Low-Cost Fabrication of Magic-Angle Spinning NMR Probeheads, Anal. Chem., 95, 10384–10389, https://doi.org/10.1021/acs.analchem.3c01323, 2023.
Roschmann, P.: Radiofrequency Penetration and Absorption in the Human Body – Limitations to High-Field Whole-Body Nuclear Magnetic Resonance Imaging, Med. Phys., 14, 922–931, https://doi.org/10.1118/1.595995, 1987.
Ryan, H., Smith, A., and Utz, M.: Structural Shimming for High-Resolution Nuclear Magnetic Resonance Spectroscopy in Lab-on-a-Chip Devices, Lab on a Chip, 14, 1678–1685, https://doi.org/10.1039/c3lc51431e, 2014.
Sangal, M., Anikeeva, M., Priese, S. C., Mattern, H., Hövener, J. B., and Speck, O.: MR based Magnetic Susceptibility Measurements of 3D Printing Materials at 3 Tesla, J. Magn. Reson. Open, 16–17, 100138, https://doi.org/10.1016/j.jmro.2023.100138, 2023.
Schenck, J. F.: The Role of Magnetic Susceptibility in Magnetic Resonance Imaging: MRI Magnetic Compatibility of the First and Second Kinds, Med. Phys., 23, 815–850, https://doi.org/10.1118/1.597854, 1996.
States, D. J., Haberkorn, R. A., and Ruben, D. J.: A Two-dimensional Nuclear Overhauser Experiment with Pure Absorpition Phase in Four Quadrants, J. Magn. Reson., 48, 286–292, https://doi.org/10.1016/0022-2364(82)90279-7, 1982.
Stott, K., Stonehouse, J., Keeler, J., Hwang, T. L., and Shaka, A. J.: Excitation Sculpting in High-Resolution Nuclear Magnetic Resonance Spectroscopy – Application to Selective NOE Experiments, J. Am. Chem. Soc., 117, 4199–4200, https://doi.org/10.1021/ja00119a048, 1995.
Terwilliger, T. C. and Eisenberg, D.: The Structure of Melittin. 2. Interpretation of the Structure, J. Biol. Chem., 257, 6016–6022, https://doi.org/10.1016/S0021-9258(20)65098-0, 1982.
Torchia, D. A.: Slight Mistuning of a Cryogenic Probe Significantly Perturbs the Water H-1 Precession Frequency, J. Biomol. NMR, 45, 241–244, https://doi.org/10.1007/s10858-009-9363-6, 2009.
Tsukada, K., Kiwa, T., and Masuda, Y.: AC Magnetic Properties of Large Volume of Water - Susceptibility Measurement in Unshielded Environment, Jap. J. Appl. Phys. Part 2, 45, L1097–L1099, https://doi.org/10.1143/jjap.45.L1097, 2006.
Ugurbil, K.: Imaging at Ultrahigh Magnetic Fields: History, Challenges, and Solutions, Neuroimage, 168, 7–32, https://doi.org/10.1016/j.neuroimage.2017.07.007, 2018.
VanderHart: Magnetic Susceptibility & High Resolution NMR of Liquids and Solids, in: Encyclopedia of Nuclear Magnetic Resonance, edited by: Grant, D. M. and Harris, R. K., Wiley, New York, https://doi.org/10.1002/9780470034590.emrstm0293, 1996.
Wapler, M. C., Leupold, J., Dragonu, J., von Elverfeld, D., Zaitsev, M., and Wallrabe, U.: Magnetic Properties of Materials for MR Engineering, Micro-MR and Beyond, J. Magn. Reson., 242, 233–242, https://doi.org/10.1016/j.jmr.2014.02.005, 2014.
Xia, Y. L., Rossi, P., Subrahmanian, M. V., Huang, C. D., Saleh, T., Olivieri, C., Kalodimos, C. G., and Veglia, G.: Enhancing the Sensitivity of Multidimensional NMR Experiments by Using Triply-Compensated π Pulses, J. Biomol. NMR, 69, 237–243, https://doi.org/10.1007/s10858-017-0153-2, 2017.
Short summary
3D-printed sample cells inserted into standard 5 mm NMR (nuclear magnetic resonance) tubes are demonstrated to offer improved sensitivity for biomolecular NMR spectroscopy when the quantity of material is limiting. Using an ellipsoidal shape, magnetic field homogeneity inside the microcell to the first order is insensitive to differences in the magnetic susceptibility of the printer resin and solvent. The sample cells are particularly useful at high ionic strength and at ultrahigh magnetic fields.
3D-printed sample cells inserted into standard 5 mm NMR (nuclear magnetic resonance) tubes are...
