Divide and Conquer: A Tailored Solid-state NMR Approach to Study Large Membrane Protein Complexes

Sheng Qi Xiang; Cecilia Pinto; Marc Baldus

doi:10.1002/anie.202203319

Divide and Conquer: A Tailored Solid-state NMR Approach to Study Large Membrane Protein Complexes

Sheng Qi Xiang^*
, Cecilia Pinto
, Marc Baldus

^*Corresponding author for this work

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

Abstract

Membrane proteins are known to exert many essential biological functions by forming complexes in cell membranes. An example refers to the β-barrel assembly machinery (BAM), a 200 kDa pentameric complex containing BAM proteins A–E that catalyzes the essential process of protein insertion into the outer membrane of gram-negative bacteria. While progress has been made in capturing three-dimensional structural snapshots of the BAM complex, the role of the lipoprotein BamC in the complex assembly in functional lipid bilayers has remained unclear. We have devised a component-selective preparation scheme to directly study BamC as part of the entire BAM complex in lipid bilayers. Combination with proton-detected solid-state NMR methods allowed us to probe the structure, dynamics, and supramolecular topology of full-length BamC embedded in the entire complex in lipid bilayers. Our approach may help decipher how individual proteins contribute to the dynamic formation and functioning of membrane protein complexes in membranes.

Original language	English
Article number	e202203319
Number of pages	6
Journal	Angewandte Chemie - International Edition
Volume	61
Issue number	33
DOIs	https://doi.org/10.1002/anie.202203319
Publication status	Published - 15 Aug 2022

Bibliographical note

Funding Information:
We thank Gert Folkers for helpful discussions, Johan van der Zwan and Mark Daniels for technical support. The plasmid pJH114 was kindly provided by Harris Bernstein et al. (NIH). This work was supported by NWO (the Dutch Science Foundation), through a VENI grant (number 722.016.002) to S.X., as well as GROOT (number 175.010.2009.002) and TOP-PUNT (number 718.015.001) grants to MB, and the National Roadmap Large-Scale NMR Facility of the Netherlands (grant number 184.032.207).

Funding Information:
We thank Gert Folkers for helpful discussions, Johan van der Zwan and Mark Daniels for technical support. The plasmid pJH114 was kindly provided by Harris Bernstein et al. (NIH). This work was supported by NWO (the Dutch Science Foundation), through a VENI grant (number 722.016.002) to S.X., as well as GROOT (number 175.010.2009.002) and TOP‐PUNT (number 718.015.001) grants to MB, and the National Roadmap Large‐Scale NMR Facility of the Netherlands (grant number 184.032.207).

Publisher Copyright:
© 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.

Funding

We thank Gert Folkers for helpful discussions, Johan van der Zwan and Mark Daniels for technical support. The plasmid pJH114 was kindly provided by Harris Bernstein et al. (NIH). This work was supported by NWO (the Dutch Science Foundation), through a VENI grant (number 722.016.002) to S.X., as well as GROOT (number 175.010.2009.002) and TOP-PUNT (number 718.015.001) grants to MB, and the National Roadmap Large-Scale NMR Facility of the Netherlands (grant number 184.032.207). We thank Gert Folkers for helpful discussions, Johan van der Zwan and Mark Daniels for technical support. The plasmid pJH114 was kindly provided by Harris Bernstein et al. (NIH). This work was supported by NWO (the Dutch Science Foundation), through a VENI grant (number 722.016.002) to S.X., as well as GROOT (number 175.010.2009.002) and TOP‐PUNT (number 718.015.001) grants to MB, and the National Roadmap Large‐Scale NMR Facility of the Netherlands (grant number 184.032.207).

Keywords

BAM Complex
Isotopic Labelling
Membrane Protein Complex
NMR Spectroscopy
Proton Detection

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Divide and ConquerFinal published version, 3.56 MBLicence: CC BY

Cite this

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title = "Divide and Conquer: A Tailored Solid-state NMR Approach to Study Large Membrane Protein Complexes",

abstract = "Membrane proteins are known to exert many essential biological functions by forming complexes in cell membranes. An example refers to the β-barrel assembly machinery (BAM), a 200 kDa pentameric complex containing BAM proteins A–E that catalyzes the essential process of protein insertion into the outer membrane of gram-negative bacteria. While progress has been made in capturing three-dimensional structural snapshots of the BAM complex, the role of the lipoprotein BamC in the complex assembly in functional lipid bilayers has remained unclear. We have devised a component-selective preparation scheme to directly study BamC as part of the entire BAM complex in lipid bilayers. Combination with proton-detected solid-state NMR methods allowed us to probe the structure, dynamics, and supramolecular topology of full-length BamC embedded in the entire complex in lipid bilayers. Our approach may help decipher how individual proteins contribute to the dynamic formation and functioning of membrane protein complexes in membranes.",

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note = "Funding Information: We thank Gert Folkers for helpful discussions, Johan van der Zwan and Mark Daniels for technical support. The plasmid pJH114 was kindly provided by Harris Bernstein et al. (NIH). This work was supported by NWO (the Dutch Science Foundation), through a VENI grant (number 722.016.002) to S.X., as well as GROOT (number 175.010.2009.002) and TOP-PUNT (number 718.015.001) grants to MB, and the National Roadmap Large-Scale NMR Facility of the Netherlands (grant number 184.032.207). Funding Information: We thank Gert Folkers for helpful discussions, Johan van der Zwan and Mark Daniels for technical support. The plasmid pJH114 was kindly provided by Harris Bernstein et al. (NIH). This work was supported by NWO (the Dutch Science Foundation), through a VENI grant (number 722.016.002) to S.X., as well as GROOT (number 175.010.2009.002) and TOP‐PUNT (number 718.015.001) grants to MB, and the National Roadmap Large‐Scale NMR Facility of the Netherlands (grant number 184.032.207). Publisher Copyright: {\textcopyright} 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.",

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N2 - Membrane proteins are known to exert many essential biological functions by forming complexes in cell membranes. An example refers to the β-barrel assembly machinery (BAM), a 200 kDa pentameric complex containing BAM proteins A–E that catalyzes the essential process of protein insertion into the outer membrane of gram-negative bacteria. While progress has been made in capturing three-dimensional structural snapshots of the BAM complex, the role of the lipoprotein BamC in the complex assembly in functional lipid bilayers has remained unclear. We have devised a component-selective preparation scheme to directly study BamC as part of the entire BAM complex in lipid bilayers. Combination with proton-detected solid-state NMR methods allowed us to probe the structure, dynamics, and supramolecular topology of full-length BamC embedded in the entire complex in lipid bilayers. Our approach may help decipher how individual proteins contribute to the dynamic formation and functioning of membrane protein complexes in membranes.

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KW - BAM Complex

KW - Isotopic Labelling

KW - Membrane Protein Complex

KW - NMR Spectroscopy

KW - Proton Detection

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DO - 10.1002/anie.202203319

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JO - Angewandte Chemie - International Edition

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IS - 33

M1 - e202203319

ER -

Divide and Conquer: A Tailored Solid-state NMR Approach to Study Large Membrane Protein Complexes

Abstract

Bibliographical note

Funding

Keywords

Access to Document

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