ASILOMAR, California – April 20, 2015 – At the 56th Experimental Nuclear Magnetic Resonance Conference (www.enc-conference.org), Bruker this week is launching its next-generation of GHz-class NMR technology, with a combination of major method and instrumentation advances, which enable even more advanced scientific and translational research in structural biology, drug discovery and the study of macromolecular complexes.
The primary focus of Bruker’s unique, next-generation GigaHertz (GHz) Nuclear Magnetic Resonance (NMR) spectroscopy technology is to enable breakthrough fundamental research in molecular and cell biology on Intrinsically Disordered Proteins (IDPs). In particular, ultra-high field NMR in combination with other experimental and computational methods, has recently been shown to enable more and more detailed studies of the structural ensembles, post-translational modifications, dynamics, multiple interactions, specific binding partners, signaling and regulatory roles, formation of membrane-less cellular organelles, and other important functions of Intrinsically Disordered Proteins (IDPs). Due to the scarcity of understanding of the molecular functions for the vast majority of IDPs, they are sometimes also referred to as the ‘Dark Proteome’. Please note the link to a short video on IDPs below.
The next-generation GHz NMR technology is the result of a confluence of recent break-through scientific discoveries, major technical achievements and key, customer-driven new NMR methods development, including:
- New Actively-Shielded 1 GHz NMR Magnets
- Novel High-Dimensionality and Fast Acquisition NMR Methods
- 13C and Novel 15N Direct Detection for Large Proteins and IDPs
- Advanced Parallel NMR with Multiple Receiver Acquisition
- New 3 mm TCI CryoProbe for GHz-class Indirect Experiments
- New Triple-Gradient 5mm CryoProbes
- Novel Single-Story Ascend Aeon 900 MHz Magnets
- New 1 GHz ultra-fast 111 kHz MAS solid-state NMR probe (see separate press release)
For the scientific community, and scientific press and media, a Scientific and Technical Section is provided below.
Frank H. Laukien, Ph.D., President and CEO of Bruker Corporation, commented: “The study of IDPs is one of the most important next frontiers in biology and in understanding disease pathogenesis. We are very excited to usher in the next-generation of GHz NMR technology. Its primary mission is to enable molecular and cell biologists to accelerate their quest to illuminate the ‘Dark Proteome’, with expected enormous benefits for healthcare and patients. The next-generation GHz NMR tools for IDP research are expected to dramatically accelerate our understanding of many fundamental biological processes. Moreover, IDP research has already delivered key discoveries, and offers great promise, for breakthroughs in the study of cancer biology and neurodegenerative diseases, like Alzheimer’s.”
Actively Shielded Aeon 1 GHz NMR Magnet
A Bruker 1 GHz NMR system with a first-generation, unshielded 23.5 Tesla magnet has been running very successfully at the Ultra-High Field NMR Center in Lyon, France since 2009, with remarkable scientific output.
Bruker intends to deliver the world’s first, next-generation 1GHz NMR systems with actively-shielded Aeon™ 1 GHz magnets to the University of Bayreuth, Germany, in late 2015, and to the University of Toronto, Canada in the first half of 2016. Active shielding reduces the space volume occupied by the 5 Gauss stray field of this two-story magnet by more than one order of magnitude, and makes siting of next-generation Aeon™ 1 GHz magnets straight-forward.
The new Aeon™ 1 GHz magnets have been developed using the latest, proprietary, advanced superconductors from Bruker’s Energy & Supercon Technologies (BEST) division. The Aeon™ 1 GHz also features proprietary, fully-integrated, novel, active refrigeration technology, which eliminates the need for liquid Nitrogen completely, and brings liquid Helium boil-off essentially to zero in normal operation. Bi-annual pulse-tube cooler maintenance can be done at full field with minimal disruption and down-time. By mid-2016, Bruker expects to have capacity for four to six Aeon™ 1 GHz magnets annually.
Recent years have seen breakthroughs developed by the NMR research community in APSY (automated projection spectroscopy) and in fast NMR acquisition using Non-Uniform Sampling (NUS). These and other seminal NMR methods, pulse programming and data analysis advances, together with GHz-class NMR sensitivity and spectral dispersion, are essential for increasingly automated, sequence-specific backbone and side chain resonance assignments of larger globular proteins, and their complexes, from chemical shift correlations determined by nD (n >= 4) NMR experiments. Fast high-dimensional methods like NUS, projection spectroscopy and APSY methods are particularly needed for IDPs, or proteins with long intrinsically disordered regions (IDRs) due to the inherently lower spectral dispersion of many IDPs/IDRs.
Bruker’s latest NMR software version TopSpin™ 3.5 now enables projection spectroscopy, including APSY-NMR, and routine NUS acquisition, and supports automatic execution of suitable pulse programs with fast acquisition techniques, with access to up to 6-dimensional NMR experiments at significantly reduced acquisition times. NUS techniques acquire only a subset of the data points of high dimensionality experiments and use novel reconstruction methods that ultimately allow the extraction of complete sets of chemical shift information. One new introduction in the field of reconstruction of non-uniformly sampled data sets is compressed sensing (CS) . It provides a general approach for a major reduction of measuring time and quality improvement of the sparsely detected spectra.
These recent and now fully integrated NMR techniques are taking full advantage of the advances in GHz NMR sensitivity, due to highest magnetic field strengths and new CryoProbe developments (see below), in cases where the time required to acquire high-dimensionality data set in conventional ways would be completely prohibitive. The confluence of major strides in NMR methodology, software and NMR technology together have enabled the next-generation level of break-through NMR performance that is a prerequisite for large-scale functional and disease-directed exploration of the universe of IDPs/IDRs, which today remains largely unexplored.
Professor Vladislav Orekhov from the Swedish NMR Center at the University of Gothenburg said: “I am fascinated by the rapid progress in the theory and applications of novel signal acquisition and processing techniques that push boundaries of the dimensionality and resolution of NMR spectra, and permit real-time investigation of biological processes with atomic resolution. A combination of ultra-high field GHz magnets and non-uniform sampling techniques greatly enhances our capability to tackle challenging biomedical problems, including the characterization of large protein machines and intrinsically disordered proteins. I have no doubts we are witnessing the most striking development in this field!”
Direct 13C detection of isotopically labelled proteins has become an essential tool in high-field NMR in recent years, with well-known advantages for metalloproteins and IDPs, and providing complementary information to main-stream indirect 1H-detected NMR experiments in structural biology. The introduction of next-generation 5mm TXO and 5mm TCI CryoProbes up to 1 GHz, both using cold 15N preamplifiers, in conjunction with the latest GHz-class magnets, enabled by recent, novel 15N direct-detect NMR methods, now make direct 15N detection advantageous, sensitive and quite useful in very large globular proteins and in IDPs, due to the longer relaxation times, high resolution and low chemical shift anisotropy of 15N spectra.
Furthermore, 15N detection can be beneficial in cases where carbon-detected methods suffer from multiple couplings to neighboring carbons, or in the study of proline-rich protein domains. Another attractive area of 15N detection are paramagnetic metallo-proteins, where 1H or even 13C magnetization is broadened beyond detection limits. These 15N detected experiments are critically dependent on the high sensitivity delivered by the new 5mm TXO or TCI CryoProbes with cryogenic 15N preamplifiers, and it turns out that 15N-TROSY experiments are expected to have their highest sensivitiy benefit at around 1 GHz proton frequency.
Professor Gerhard Wagner of Harvard Medical School, a pioneer of 15N direct detection, stated: “Direct 15N and 13C detection methods have recently been evolved and found to be almost as sensitive as 1H detection techniques, benefitting from the slow transverse relaxation. This opens new opportunities for studies of proline-rich polypeptides often found in regulatory regions, such as phosphorylation domains. However, spectra of such domains are typically poorly dispersed and highest field strengths will be needed to reveal mechanisms of phosphorylation-dependent switch mechanisms. Availability of GHz-class NMR instruments will be important for revealing mechanisms of phosphorylation-dependent signaling switches.”
The unique multiple receiver technology of Bruker’s AVANCE III HD platform now enables elegant and efficient polarization sharing and parallel acquisition NMR spectroscopy to detect simultaneously signals from multiple nuclear species, such as 1H, 2D, 13C, 15N, 19F and 31P. The multi-receiver experiments can also be used in combination with the fast acquisition schemes such as projection-reconstruction techniques and can provide significantly more information from a single NMR measurement, compared to the conventional single receiver techniques. Parallel NMR with multi-receiver acquisition is also well suited for structure verification or elucidation of small molecules in drug development and discovery, as well as for higher dimensionality experiments for studying globular proteins or IDPs.
Professor Markus Zweckstetter, Group Leader at the German Center for Neurodegenerative Diseases at the University of Göttingen, stated: “We are thrilled by the new opportunities to use next-generation GHz NMR technology to perform 5-7 dimensional NMR experiments on intrinsically disordered proteins (IDPs) of high medical relevance, as well as solid-state NMR experiments on membrane proteins in native-like environments, or on complexes of IDPs with diagnostic and therapeutic ligands. In particular, we are excited about further boosting the resolution of 6-dimensional experiments on IDPs with GHz-class NMR spectrometers with next-generation CryoProbes and parallel NMR capabilities by a significant factor, compared to traditional high-field NMR spectrometers. We are convinced that this step-change in information content on IDP function will help the scientific community to unravel the ‘Dark Proteome’ of IDPs, and enable completely new insights into key biological and disease processes, particularly in cancer and neurodegenerative diseases.”
This new design of smaller diameter NMR CryoProbes is advantageous for highest-field GHz biomolecular applications, when sample volumes are limited and sample salt concentrations are in the physiological range. The new 3mm design also delivers shorter radiofrequency (RF) pulses at equivalent RF power, compared to 5 mm CryoProbes, an important benefit for IDP dynamics experiments, when working at GHz fields.
Professor Göran Karlsson, Director of the Swedish NMR Center at the University of Gothenburg, was Bruker’s key collaborator in the development of 3mm CryoProbes. He commented: “The performance of the new 3 mm TCI CryoProbe on our 800 MHz is just spectacular. For some time we have been working with 3mm samples in structural biology, and with the new 800 MHz 3mm CryoProbe we obtain dramatic S/N increase . In metabolomics applications, we benefit from both, the increase in S/N and the ability to work with reduced sample volumes. For many applications in life science research, the 3mm CryoProbe represents a leap forward. We’re planning to order the 3mm CryoProbe also for our 900 MHz spectrometer."
The latest 5mm TCI CryoProbes can now optionally be equipped with actively-shielded, triple-axis pulsed field gradient coils. Triple axis gradients enable faster pulse sequence optimization with respect to gradient coherence selection. In addition, the residual water signal is typically reduced by a factor of 2-3 compared to single axis gradient probes. In addition to localized spectroscopy, recently published fast methods such as SMART NMR also become accessible.
Professor Lewis Kay at the University of Toronto, Canada, was Bruker’s key collaborator in the development of triple-axis gradient CryoProbes. Dr. Kay explained: “We are eagerly anticipating delivery of triple-axis gradient CryoProbes for our 600 MHz, 800 MHz and 1 GHz spectrometers. The ability to significantly improve water suppression by replacing Z-coherence transfer selection gradients with those along X or Y, for example, makes optimum pulse sequence development easier. Better suppression of water will lead to significantly less noise, especially near the water line, as well as better baselines, translating into higher quality data sets and subsequent improvement in signal to noise.”
Single-Story Ascend Aeon 900 MHz Magnet
The world’s first compact, single-story 900 MHz NMR magnet for high-resolution protein NMR which is being introduced at ENC 2015, integrates advanced refrigeration technology, and obviates the need for any cryogen refills. Previous 900 MHz magnets required two-story laboratories, limiting the wider adoption of ultra-high field NMR, except in specialized NMR laboratories.
Professor Paul Rösch is the Director of the Research Center for Bio-Macromolecules at the University of Bayreuth, Germany, where the installation of the world’s first Ascend Aeon 900 magnet for high-resolution NMR was recently completed. Dr. Rösch stated: “Our new Ascend Aeon 900 magnet enables long-term, helium consumption-free operation without user maintenance. The reduced height and stray fields of this novel, compact, ultra-high field magnet, maximize siting flexibility and thus reduce laboratory space costs. From our perspective both factors are key requirements to further grow the adoption of ultra-high field NMR in biology, and also to expand into clinical research. We’re very pleased with the stability and the performance of our new Ascend Aeon 900 magnet.”
The University of Bayreuth will also be the site for the world’s first installation of a shielded 1 GHz magnet, presently expected in the fourth quarter of 2015.
For more than 50 years, Bruker has enabled scientists to make breakthrough discoveries and develop new applications that improve the quality of human life. Bruker’s high-performance scientific research instruments and high-value analytical solutions enable scientists to explore life and materials at molecular, cellular and microscopic levels.
In close cooperation with our customers, Bruker is enabling innovation, productivity and customer success in life science molecular research, in applied and pharma applications, in microscopy, nano-analysis and industrial applications, as well as in cell biology, preclinical imaging, clinical research, microbiology and molecular diagnostics.
For more information on Bruker at ENC 2015, please visit www.bruker.com/enc
For a short, educational video on IDPs, please go to: www.idpbynmr.eu/home/video.html
Dr. Thorsten Thiel
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