High-resolution studies of proteins in natural membranes by solid-state nmr

David Beriashvili; Raymond D. Schellevis; Federico Napoli; Markus Weingarth; Marc Baldus

doi:10.3791/62197

High-resolution studies of proteins in natural membranes by solid-state nmr

David Beriashvili
, Raymond D. Schellevis
, Federico Napoli
, Markus Weingarth^*
, Marc Baldus

^*Corresponding author for this work

Utrecht University

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Membrane proteins are vital for cell function and thus represent important drug targets. Solid-state Nuclear Magnetic Resonance (ssNMR) spectroscopy offers a unique access to probe the structure and dynamics of such proteins in biological membranes of increasing complexity. Here, we present modern solid-state NMR spectroscopy as a tool to study structure and dynamics of proteins in natural lipid membranes and at atomic scale. Such spectroscopic studies profit from the use of high-sensitivity ssNMR methods, i.e., proton-(¹ H)-detected ssNMR and DNP (Dynamic Nuclear Polarization) supported ssNMR. Using bacterial outer membrane beta-barrel protein BamA and the ion channel KcsA, we present methods to prepare isotope-labeled membrane proteins and to derive structural and motional information by ssNMR.

Original language	English
Article number	e62197
Pages (from-to)	1-22
Number of pages	22
Journal	Journal of Visualized Experiments
Volume	2021
Issue number	169
DOIs	https://doi.org/10.3791/62197
Publication status	Published - 3 Mar 2021

Bibliographical note

Funding Information:
This work is part of the research programs ECHO, TOP, TOP-PUNT, VICI, and VIDI with project numbers 723.014.003, 711.018.001, 700.26.121, 700.10.443, and 718.015.00, which are financed by the Dutch Research Council (NWO).

Publisher Copyright:
© 2021 JoVE Journal of Visualized Experiments.

Funding

This work is part of the research programs ECHO, TOP, TOP-PUNT, VICI, and VIDI with project numbers 723.014.003, 711.018.001, 700.26.121, 700.10.443, and 718.015.00, which are financed by the Dutch Research Council (NWO).

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jove-protocol-62197-highresolution-studies-of-proteins-in-natural-membranes-by-solidstate-nmrFinal published version, 1.12 MBLicence: Taverne

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title = "High-resolution studies of proteins in natural membranes by solid-state nmr",

abstract = "Membrane proteins are vital for cell function and thus represent important drug targets. Solid-state Nuclear Magnetic Resonance (ssNMR) spectroscopy offers a unique access to probe the structure and dynamics of such proteins in biological membranes of increasing complexity. Here, we present modern solid-state NMR spectroscopy as a tool to study structure and dynamics of proteins in natural lipid membranes and at atomic scale. Such spectroscopic studies profit from the use of high-sensitivity ssNMR methods, i.e., proton-(1 H)-detected ssNMR and DNP (Dynamic Nuclear Polarization) supported ssNMR. Using bacterial outer membrane beta-barrel protein BamA and the ion channel KcsA, we present methods to prepare isotope-labeled membrane proteins and to derive structural and motional information by ssNMR.",

author = "David Beriashvili and Schellevis, \{Raymond D.\} and Federico Napoli and Markus Weingarth and Marc Baldus",

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doi = "10.3791/62197",

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AU - Beriashvili, David

AU - Schellevis, Raymond D.

AU - Napoli, Federico

AU - Weingarth, Markus

AU - Baldus, Marc

N1 - Funding Information: This work is part of the research programs ECHO, TOP, TOP-PUNT, VICI, and VIDI with project numbers 723.014.003, 711.018.001, 700.26.121, 700.10.443, and 718.015.00, which are financed by the Dutch Research Council (NWO). Publisher Copyright: © 2021 JoVE Journal of Visualized Experiments.

PY - 2021/3/3

Y1 - 2021/3/3

N2 - Membrane proteins are vital for cell function and thus represent important drug targets. Solid-state Nuclear Magnetic Resonance (ssNMR) spectroscopy offers a unique access to probe the structure and dynamics of such proteins in biological membranes of increasing complexity. Here, we present modern solid-state NMR spectroscopy as a tool to study structure and dynamics of proteins in natural lipid membranes and at atomic scale. Such spectroscopic studies profit from the use of high-sensitivity ssNMR methods, i.e., proton-(1 H)-detected ssNMR and DNP (Dynamic Nuclear Polarization) supported ssNMR. Using bacterial outer membrane beta-barrel protein BamA and the ion channel KcsA, we present methods to prepare isotope-labeled membrane proteins and to derive structural and motional information by ssNMR.

AB - Membrane proteins are vital for cell function and thus represent important drug targets. Solid-state Nuclear Magnetic Resonance (ssNMR) spectroscopy offers a unique access to probe the structure and dynamics of such proteins in biological membranes of increasing complexity. Here, we present modern solid-state NMR spectroscopy as a tool to study structure and dynamics of proteins in natural lipid membranes and at atomic scale. Such spectroscopic studies profit from the use of high-sensitivity ssNMR methods, i.e., proton-(1 H)-detected ssNMR and DNP (Dynamic Nuclear Polarization) supported ssNMR. Using bacterial outer membrane beta-barrel protein BamA and the ion channel KcsA, we present methods to prepare isotope-labeled membrane proteins and to derive structural and motional information by ssNMR.

UR - https://www.scopus.com/pages/publications/85102586824

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DO - 10.3791/62197

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SN - 1940-087X

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JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

IS - 169

M1 - e62197

ER -

High-resolution studies of proteins in natural membranes by solid-state nmr

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