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Yang L, Catalano C, Xu Y, et al. A native cell membrane nanoparticles system allows for high-quality functional proteoliposome reconstitution. BBA Adv. 2021;1doi: 10.1016/j.bbadva.2021.100011.
Yang, L., Catalano, C., Xu, Y., Qiu, W., Zhang, D., McDermott, A., Guo, Y., & Blount, P. (2021). A native cell membrane nanoparticles system allows for high-quality functional proteoliposome reconstitution. BBA advances, 1. https://doi.org/10.1016/j.bbadva.2021.100011
Yang, Limin, et al. "A native cell membrane nanoparticles system allows for high-quality functional proteoliposome reconstitution." BBA advances vol. 1 (2021). doi: https://doi.org/10.1016/j.bbadva.2021.100011
Yang L, Catalano C, Xu Y, Qiu W, Zhang D, McDermott A, Guo Y, Blount P. A native cell membrane nanoparticles system allows for high-quality functional proteoliposome reconstitution. BBA Adv. 2021;1. doi: 10.1016/j.bbadva.2021.100011. Epub 2021 Apr 05. PMID: 34296205; PMCID: PMC8294337.
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BBA Adv. 2021;1. doi: 10.1016/j.bbadva.2021.100011. Epub 2021 Apr 05.

A native cell membrane nanoparticles system allows for high-quality functional proteoliposome reconstitution.

BBA advances

Limin Yang, Claudio Catalano, Yunyao Xu, Weihua Qiu, Dongyu Zhang, Ann McDermott, Youzhong Guo, Paul Blount

Affiliations

  1. Department of Physiology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9040, United States.
  2. Department of Medicinal Chemistry, Virginia Commonwealth University, Richmond, VA 23298-0540, United States.
  3. Institute for Structural Biology, Drug Discovery and Development, Virginia Commonwealth University, Richmond, VA 23298-0133, United States.
  4. Department of Chemistry, Columbia University, New York, NY 10027, United States.

PMID: 34296205 PMCID: PMC8294337 DOI: 10.1016/j.bbadva.2021.100011

Abstract

Proteoliposomes mimic the cell membrane environment allowing for structural and functional membrane protein analyses as well as antigen presenting and drug delivery devices. To make proteoliposomes, purified functional membrane proteins are required. Detergents have traditionally been used for the first step in this process However, they can irreversibly denature or render membrane proteins unstable, and the necessary removal of detergents after reconstitution can decrease proteoliposome yields. The recently developed native cell membrane nanoparticles (NCMN) system has provided a variety of detergent-free alternatives for membrane protein preparation for structural biology research. Here we attempt to employ the MCMN system for the functional reconstitution of channels into proteoliposomes. NCMN polymers NCMNP1-1 and NCMNP7-1, members of a NCMN polymer library that have been successful in extraction and affinity purification of a number of intrinsic membrane proteins, were selected for the purification and subsequent reconstitution of three bacterial channels: KcsA and the mechanosensitive channels of large and small conductance (MscL and MscS). We found that channels in NCMN particles, which appeared to be remarkably stable when stored at 4 °C, can be reconstituted into bilayers by simply incubating with lipids. We show that the resulting proteoliposomes can be patched for electrophysiological studies or used for the generation of liposome-based nanodevices. In sum, the findings demonstrate that the NCMN system is a simple and robust membrane protein extraction and reconstitution approach for making high-quality functional proteoliposomes that could significantly impact membrane protein research and the development of nanodevices.

Keywords: KcsA; MscL; MscS; NCMN; Proteoliposome; Triggered-release

Conflict of interest statement

Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this pa

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