Rebecca Calamandrei (CERM, Italy)
Abstract: The ITACA.SB project (https://www.itaca-sb.it/about/) is dedicated to potentiate the Italian Instruct-ERIC center, CERM/CIRMMP (https://www.cerm.unifi.it/) and significantly enhance structural biology (SB) services at selected laboratories of CNR. By enhancing service capacity and overcoming key access barriers, the project supports high-level life sciences research in Italy, boosts international visibility, and fosters stronger integration with European research infrastructures.
Within this embodiment, a significant enhancement of the instrumentation at CERM/CIRMMP has enabled the expansion of both solution and solid-state NMR research as well as the biotechnologies instrumentation ranging a broad spectrum of experiment set-up and characterization techniques.
As part of the infrastructure upgrades supported by ITACA.SB, the 1.2 GHz NMR spectrometer at CERM has been equipped with a 0.7 mm solid-state MAS probe. This high-field system offers exceptional performance for the investigation of solid-phase materials, including protein crystals and, more critically, non-crystalline systems such as amyloid fibrils, membrane proteins, and complex sediments. The implementation of ultra-fast magic angle spinning at 1.2 GHz enables the acquisition of high-resolution, proton-detected spectra, comparable in quality to those obtained in solution-state NMR. This advancement significantly expands the capabilities of solid-state NMR for probing molecular dynamics and intermolecular interactions in challenging biological and material samples.
As the result of the synergic integration of upgraded infrastructure, targeted user support, and strategic collaboration, ITACA.SB not only strengthens Italy’s contribution to the structural biology landscape but also ensures that CERM/CIRMMP operates as a competitive hub for research, facilitating the alignment within the European Research area.
-
Hello Rebecca! Nice presentation!
Among all these beautiful instruments and applications presented here, I was intrigued by the performances of the 1.2 GHz equipped with the 0.7 mm MAS probe.
Could you please comment about the resolution that can be obtained? How fast can you spin and what nuclear spins can be detected?-
Hi Marco,
Thanks for your comment! The 0.7 mm MAS probe is capable of spinning up to 111 kHz and features three channels dedicated to the detection of ¹H, ¹³C, and ¹⁵N. The major benefits of using this type of probe at ultra-high magnetic fields are particularly evident in ¹H-detected spectra, which can achieve a level of resolution comparable to that of solution-state NMR. This enables detailed studies of residue-specific dynamics and protein–ligand interactions. These findings highlight the crucial importance of combining ultra-high magnetic fields with ultra-fast magic angle spinning for the structural and dynamic characterization of biomolecular systems in the solid state.
-
-
Nice presentation. I am curious about the NEO console. Does this mean one can set up two different experiments and both run simultaneously instead of queuing experiments?
-
Thank you for your kind and relevant question. In the novel NEO console, each radiofrequency (RF) channel is equipped with both transmission and reception capabilities. This design effectively allows each channel to operate as an independent spectrometer, with its own RF generation, transmission, and receiver architecture.
In practice, this enables the implementation of multi-receiver experiments in a user-friendly way. The multiple receiver approach developed at our research infrastructure exploits the recovery delay of one experiment to acquire additional experiments simultaneously (see: [Biophys. J. 2019, 10.1016/j.bpj.2019.05.017]).For example, ¹³C- and ¹H-detected experiments can be combined to obtain complementary information on multidomain proteins ([Biomolecules 2022, 10.3390/biom12070929]) or to monitor complex protein–protein interactions in real time ([J. Am. Chem. Soc. 2024, 10.1021/jacs.4c09176]).
This simultaneous acquisition strategy is a key advantage of the NEO architecture, going beyond traditional queuing of experiments.
-
-
Nice presentation of the facility!
Is there any plans to use the 1.2 GHz with a liquid state probe?
Smaller rotor means less materials. How does the sensitivity of the 0.7mm rotor compares with 1.3/1.9 mm rotors on a GHz for example?-
Dear Nicolas Bolik-Coulon,
Thank you for your question. Indeed, we have a 5 mm CP-TXO probe for 13C direct detection that is also routinely used at the 1.2 GHz instrument. The gain in resolution at ultra-high fields is significant not only for solid-state but also for solution-state NMR experiments. This is particularly beneficial when working with biomolecules whose spectra display extensive peak overlap. The combination of ultra-high field and 13C detection helps to partially overcome the spectral crowding typically observed in IDPs and IDRs, as demonstrated in this study: [doi: 10.1038/s41596-023-00921-9]. Moreover, although the amount of sample decreases when moving from larger to smaller rotors, the linewidth also narrows due to a greater averaging of dipolar couplings, resulting in more intense signals. Additionally, the sensitivity loss caused by the reduced sample volume is partially compensated by improved inductive coupling between the coil and the sample, which becomes more efficient as the coil size decreases, as illustrated in this review [doi:10.1021/acs.chemrev.1c00918].
-
Leave a Reply