Optimizing water hyperpolarization and dissolution for sensitivity-enhanced 2D biomolecular NMR

Olsen, Gregory L.; Markhasin, Evgeny; Szekely, Or; Bretschneider, Christian O.; Frydman, Lucio

doi:10.1016/j.jmr.2016.01.005

Cited by 48 publications

(67 citation statements)

References 43 publications

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“…Finally, a SOFAST‐HMQC sequence composed of 32 increments in the indirect dimension, using 1 scan per increment and gradient selection (for 15 N and 1 H) could be recorded on the hyperpolarized protein by using the approach of Frydman and co‐workers . The concentration of OPN was 1.5 m m in 150 μL PBS buffer before injection of 750 μL of the hyperpolarized solution containing HDO (Figure ).…”

Section: Resultsmentioning

confidence: 99%

“…Strategies have been proposed for neutralizing the polarizing agents via precipitation and filtration, solvent extraction, or scavenging, which typically require a few seconds during which relaxation occurs. We have recently introduced a new class of polarizing agents called HYPSO (HYbrid Polarizing SOlids) consisting of mesoporous silica matrices with free radicals covalently anchored at the surface of the porous network.…”

Section: Introductionmentioning

confidence: 99%

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Filterable Agents for Hyperpolarization of Water, Metabolites, and Proteins

Vuichoud

Bornet

Nanteuil

et al. 2016

Chemistry A European J

105

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Hyperpolarization is generated by dissolution dynamic nuclear polarization (d-DNP) using a polymer-based polarizing agent dubbed FLAP (filterable labeled agents for polarization). It consists of a thermo-responsive poly(N-isopropylacrylamide), also known as pNiPAM-COOH, labeled with nitroxide radicals. The polymer powder is impregnated with an arbitrary solution of interest and frozen as is. Dissolution is followed by a simple filtration, leading to hyperpolarized solutions free from any contaminants. We demonstrated the use of FLAP to hyperpolarize partially deuterated water up to P((1) H)=6 % with a long relaxation T1 >36 s characteristic of high purity. Water hyperpolarization can be transferred to drugs, metabolites, or proteins that are waiting in an NMR spectrometer, either by exchange of labile protons or through intermolecular Overhauser effects. We also show that FLAPs are suitable polarizing agents for (13) C-labeled metabolites such as pyruvate, acetate, and alanine.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Filterable Agents for Hyperpolarization of Water, Metabolites, and Proteins

Vuichoud

Bornet

Nanteuil

et al. 2016

Chemistry A European J

105

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“…Polarization transfer from molecules that are directly hyperpolarized on proton spins using dissolution dynamic nuclear polarization (Ardenkjær-Larsen et al 2003) (D-DNP) is also possible. Ultrafast 2D NMR spectra of polarization transferred from water to exchangeable amide protons in proteins were reported (Olsen et al 2016). In our previous work, we have demonstrated that polarization transfer from a specifically binding hyperpolarized ligand shows a spectrum similar to the frequency dependence of the transfer of saturation from the protein to the ligand (Min et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Site specific polarization transfer from a hyperpolarized ligand of dihydrofolate reductase

2016

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Protein–ligand interaction is often characterized using polarization transfer by the intermolecular nuclear Overhauser effect (NOE). For such NOE experiments, hyperpolarization of nuclear spins presents the opportunity to increase the spin magnetization, which is transferred, by several orders of magnitude. Here, folic acid, a ligand of dihydrofolate reductase (DHFR), was hyperpolarized on 1H spins using dissolution dynamic nuclear polarization (D-DNP). Mixing hyperpolarized ligand with protein resulted in observable increases in protein 1H signal predominantly in the methyl group region of the spectra. Using 13C single quantum selection in a series of one-dimensional spectra, the carbon chemical shift ranges of the corresponding methyl groups can be elucidated. Signals observed in these hyperpolarized spectra could be confirmed using 3D isotope filtered NOESY spectra, although the hyperpolarized spectra were obtained in single scans. By further correlating the signal intensities observed in the D-DNP experiments with the occurrence of short distances in the crystal structure of the protein–ligand complex, the observed methyl proton signals could be matched to the chemical shifts of six amino acids in the active site of DHFR-folic acid binary complex. These data demonstrate that 13C chemical shift selection of protein resonances, combined with the intrinsic selectivity towards magnetization originating from the initially hyperpolarized spins, can be used for site specific characterization of protein–ligand interactions.

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“…In dissolution DNP, the sample is hyperpolarized at very low temperature and then dissolved through a stream of high‐pressure hot water , . The hyperpolarized sample can be used for a number of applications, from the enhancement of low‐abundance biomolecular systems, to chemical reactions, to metabolism in vivo , …”

Section: Biomedical Applicationsmentioning

confidence: 99%

NMR Spectroscopy and Metal Ions in Life Sciences

et al. 2018

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Metal ions play important structural and functional roles in many biological systems. The NMR spectroscopic characterization of biomolecules not only had to deal with the need of taking them into account, but also successfully exploited the characteristics of the paramagnetic metals to obtain structural information. The latter proved so important that diamagnetic proteins are in several cases made paramagnetic through substitution of a diamagnetic metal ion with a paramagnetic one or attachment of a paramagnetic tag. NMR studies of metal ions are also important for biomedical applications. Examples are provided here in the field of drug discovery and the development of contrast agents for MRI. More recently, paramagnetic metals proved useful also for improving the NMR sensitivity by dynamic nuclear polarization. Finally, NMR metabolomics studies are emerging for the identification and quantification of metal ions present in biological fluids.

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Optimizing water hyperpolarization and dissolution for sensitivity-enhanced 2D biomolecular NMR

Cited by 48 publications

References 43 publications

Filterable Agents for Hyperpolarization of Water, Metabolites, and Proteins

Filterable Agents for Hyperpolarization of Water, Metabolites, and Proteins

Site specific polarization transfer from a hyperpolarized ligand of dihydrofolate reductase

NMR Spectroscopy and Metal Ions in Life Sciences

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