A biradical-tagged phospholipid as a polarizing agent for solid-state MAS Dynamic Nuclear Polarization NMR of membrane proteins

Good, Daryl B.; Voinov, Maxim A.; Bolton, David; Ward, Meaghan E.; Sergeyev, Ivan V.; Caporini, Marc; Scheffer, Peter; Lo, Andy; Rosay, Melanie; Marek, Antonin; Brown, Leonid S.; I Smirnov, Alex; Ladizhansky, Vlad

doi:10.1016/j.ssnmr.2019.04.003

A biradical-tagged phospholipid as a polarizing agent for solid-state MAS Dynamic Nuclear Polarization NMR of membrane proteins

Journal Article · Wed Apr 17 00:00:00 EDT 2019 · Solid State Nuclear Magnetic Resonance

DOI:https://doi.org/10.1016/j.ssnmr.2019.04.003· OSTI ID:1609382

Good, Daryl B. ^[1]; Voinov, Maxim A. ^[2]; Bolton, David ^[3]; Ward, Meaghan E. ^[3]; Sergeyev, Ivan V. ^[4]; Caporini, Marc ^[4]; Scheffer, Peter ^[3]; Lo, Andy ^[3]; Rosay, Melanie ^[4]; Marek, Antonin ^[2]; ^[3]; ^[5]; Ladizhansky, Vlad ^[6]

University of Guelph, ON (Canada); DOE/OSTI
North Carolina State University, Raleigh, NC (United States)
University of Guelph, ON (Canada)
Bruker Biospin, Billerica, MA (United States)
North Carolina State University, Raleigh, NC (United States); Bruker Biospin, Billerica, MA (United States)
University of Guelph, ON (Canada); Bruker Biospin, Billerica, MA (United States)

We report a novel Dynamic Nuclear Polarization (DNP) NMR polarizing agent ToSMTSL-PTE representing a phospholipid with a biradical TOTAPOL tethered to the polar head group has been synthesized, characterized, and employed to enhance solid-state Nuclear Magnetic Resonance (SSNMR) signal of a lipid-reconstituted integral membrane protein proteorhodopsin (PR). A matrix-free PR formulation for DNP improved the absolute sensitivity of NMR signal by a factor of ca. 4 compared to a conventional preparation with TOTAPOL dispersed in a glassy glycerol/water matrix. DNP enhancements measured at 400 MHz/263 GHz and 600 MHz/395 GHz showed a strong field dependence but remained moderate at both fields, and comparable to those obtained for PR covalently modified with ToSMTSL. Additional continuous wave (CW) X-band electron paramagnetic resonance (EPR) experiments with ToSMTSL-PTE in solutions and in lipid bilayers revealed that an unfavorable conformational change of the linker connecting mononitroxides could be one of the reasons for moderate DNP enhancements. Further, differential scanning calorimetry (DSC) and CW EPR experiments indicated an inhomogeneous distribution and/or a possibility of a partial aggregation of ToSMTSL-PTE in DMPC:DMPA bilayers when the concentration of the polarizing agent was increased to 20 mol% to maximize the DNP enhancement. Thus, conformational changes and an inhomogeneous distribution of the lipid-based biradicals in lipid bilayers emerged as important factors to consider for further development of this matrix-free approach for DNP of membrane proteins.

View Accepted Manuscript (DOE)

Research Organization:: North Carolina State University, Raleigh, NC (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Science Foundation (NSF); Natural Sciences and Engineering Research Council of Canada (NSERC); North Carolina Biotechnology Centre (NCBC); USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: FG02-02ER15354

OSTI ID:: 1609382

Alternate ID(s):: OSTI ID: 1547486

Journal Information:: Solid State Nuclear Magnetic Resonance, Journal Name: Solid State Nuclear Magnetic Resonance Journal Issue: C Vol. 100; ISSN 0926-2040

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Solid-state NMR spectroscopy based atomistic view of a membrane protein unfolding pathway Xiao, Peng; Bolton, David; Munro, Rachel A. Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-019-11849-8	journal	August 2019

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
71 CLASSICAL AND QUANTUM MECHANICS
GENERAL PHYSICS
bilayer
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dynamic nuclear polarization
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signal enhancement

A biradical-tagged phospholipid as a polarizing agent for solid-state MAS Dynamic Nuclear Polarization NMR of membrane proteins

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References (51)

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