Details
| Original language | English |
|---|---|
| Article number | e202203443 |
| Journal | Chemistry - a European journal |
| Volume | 29 |
| Issue number | 16 |
| Early online date | 19 Dec 2022 |
| Publication status | Published - 16 Mar 2023 |
Abstract
Sensitivity and specificity are both crucial for the efficient solid-state NMR structure determination of large biomolecules. We present an approach that features both advantages by site-specific enhancement of NMR spectroscopic signals from the protein-RNA binding site within a ribonucleoprotein (RNP) by dynamic nuclear polarization (DNP). This approach uses modern biochemical techniques for sparse isotope labeling and exploits the molecular dynamics of 13C-labeled methyl groups exclusively present in the protein. These dynamics drive heteronuclear cross relaxation and thus allow specific hyperpolarization transfer across the biomolecular complex's interface. For the example of the L7Ae protein in complex with a 26mer guide RNA minimal construct from the box C/D complex in archaea, we demonstrate that a single methyl-nucleotide contact is responsible for most of the polarization transfer to the RNA, and that this specific transfer can be used to boost both NMR spectral sensitivity and specificity by DNP.
Keywords
- dynamic nuclear polarization, isotopic labeling, NMR spectroscopy, proteins, RNA
ASJC Scopus subject areas
- Chemical Engineering(all)
- Catalysis
- Chemistry(all)
- Organic Chemistry
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In: Chemistry - a European journal, Vol. 29, No. 16, e202203443, 16.03.2023.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Specific Signal Enhancement on an RNA-Protein Interface by Dynamic Nuclear Polarization
AU - Aladin, Victoria
AU - Sreemantula, Arun K.
AU - Biedenbänder, Thomas
AU - Marchanka, Alexander
AU - Corzilius, Björn
N1 - Funding Information: This work was funded by the Deutsche Forschungsgemeinschaft (DFG) through research grants CO 802/4‐1 and MA 5157/3‐1 and through large instrument grant INST 264/177‐1 FUGG. Further support was provided by the German Federal State of Hesse through the Center for Biomolecular Magnetic Resonance (BMRZ), Frankfurt am Main. We thank Johanna Becker‐Baldus and Clemens Glaubitz (Frankfurt) for support and access to the MAS DNP instrument. TB acknowledges support by the Joachim Herz Foundation through an Add‐on Fellowship for Interdisciplinary Life Sciences. Open Access funding enabled and organized by Projekt DEAL.
PY - 2023/3/16
Y1 - 2023/3/16
N2 - Sensitivity and specificity are both crucial for the efficient solid-state NMR structure determination of large biomolecules. We present an approach that features both advantages by site-specific enhancement of NMR spectroscopic signals from the protein-RNA binding site within a ribonucleoprotein (RNP) by dynamic nuclear polarization (DNP). This approach uses modern biochemical techniques for sparse isotope labeling and exploits the molecular dynamics of 13C-labeled methyl groups exclusively present in the protein. These dynamics drive heteronuclear cross relaxation and thus allow specific hyperpolarization transfer across the biomolecular complex's interface. For the example of the L7Ae protein in complex with a 26mer guide RNA minimal construct from the box C/D complex in archaea, we demonstrate that a single methyl-nucleotide contact is responsible for most of the polarization transfer to the RNA, and that this specific transfer can be used to boost both NMR spectral sensitivity and specificity by DNP.
AB - Sensitivity and specificity are both crucial for the efficient solid-state NMR structure determination of large biomolecules. We present an approach that features both advantages by site-specific enhancement of NMR spectroscopic signals from the protein-RNA binding site within a ribonucleoprotein (RNP) by dynamic nuclear polarization (DNP). This approach uses modern biochemical techniques for sparse isotope labeling and exploits the molecular dynamics of 13C-labeled methyl groups exclusively present in the protein. These dynamics drive heteronuclear cross relaxation and thus allow specific hyperpolarization transfer across the biomolecular complex's interface. For the example of the L7Ae protein in complex with a 26mer guide RNA minimal construct from the box C/D complex in archaea, we demonstrate that a single methyl-nucleotide contact is responsible for most of the polarization transfer to the RNA, and that this specific transfer can be used to boost both NMR spectral sensitivity and specificity by DNP.
KW - dynamic nuclear polarization
KW - isotopic labeling
KW - NMR spectroscopy
KW - proteins
KW - RNA
UR - http://www.scopus.com/inward/record.url?scp=85147440453&partnerID=8YFLogxK
U2 - 10.1002/chem.202203443
DO - 10.1002/chem.202203443
M3 - Article
C2 - 36815335
AN - SCOPUS:85147440453
VL - 29
JO - Chemistry - a European journal
JF - Chemistry - a European journal
SN - 0947-6539
IS - 16
M1 - e202203443
ER -