Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system

Tatsuya Niwa, Yoshihiro Sasaki, Eri Uemura, Shugo Nakamura, Minato Akiyama, Mitsuru Ando, Shinichi Sawada, Sada Atu Mukai, Takuya Ueda, Hideki Taguchi, Kazunari Akiyoshi

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90% of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins.

Original languageEnglish
Article number18025
JournalScientific reports
Volume5
DOIs
Publication statusPublished - 2015 Dec 15
Externally publishedYes

Fingerprint

Cell-Free System
Liposomes
Membrane Proteins
Protein Engineering
Drug Discovery
Computational Biology
Solubility
Proteins
Escherichia coli
Pharmaceutical Preparations

ASJC Scopus subject areas

  • General

Cite this

Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system. / Niwa, Tatsuya; Sasaki, Yoshihiro; Uemura, Eri; Nakamura, Shugo; Akiyama, Minato; Ando, Mitsuru; Sawada, Shinichi; Mukai, Sada Atu; Ueda, Takuya; Taguchi, Hideki; Akiyoshi, Kazunari.

In: Scientific reports, Vol. 5, 18025, 15.12.2015.

Research output: Contribution to journalArticle

Niwa, T, Sasaki, Y, Uemura, E, Nakamura, S, Akiyama, M, Ando, M, Sawada, S, Mukai, SA, Ueda, T, Taguchi, H & Akiyoshi, K 2015, 'Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system', Scientific reports, vol. 5, 18025. https://doi.org/10.1038/srep18025
Niwa, Tatsuya ; Sasaki, Yoshihiro ; Uemura, Eri ; Nakamura, Shugo ; Akiyama, Minato ; Ando, Mitsuru ; Sawada, Shinichi ; Mukai, Sada Atu ; Ueda, Takuya ; Taguchi, Hideki ; Akiyoshi, Kazunari. / Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system. In: Scientific reports. 2015 ; Vol. 5.
@article{93a6b056ab054eb89ff10115d277c199,
title = "Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system",
abstract = "Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90{\%} of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins.",
author = "Tatsuya Niwa and Yoshihiro Sasaki and Eri Uemura and Shugo Nakamura and Minato Akiyama and Mitsuru Ando and Shinichi Sawada and Mukai, {Sada Atu} and Takuya Ueda and Hideki Taguchi and Kazunari Akiyoshi",
year = "2015",
month = "12",
day = "15",
doi = "10.1038/srep18025",
language = "English",
volume = "5",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system

AU - Niwa, Tatsuya

AU - Sasaki, Yoshihiro

AU - Uemura, Eri

AU - Nakamura, Shugo

AU - Akiyama, Minato

AU - Ando, Mitsuru

AU - Sawada, Shinichi

AU - Mukai, Sada Atu

AU - Ueda, Takuya

AU - Taguchi, Hideki

AU - Akiyoshi, Kazunari

PY - 2015/12/15

Y1 - 2015/12/15

N2 - Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90% of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins.

AB - Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90% of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins.

UR - http://www.scopus.com/inward/record.url?scp=84949765214&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84949765214&partnerID=8YFLogxK

U2 - 10.1038/srep18025

DO - 10.1038/srep18025

M3 - Article

VL - 5

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 18025

ER -