Publications

A03 KURUMA, Yutetsu |Proposed Research Projects (2014-2015)

Paper | Original Paper

2015

*Yutetsu Kuruma and Takuya Ueda,
The PURE system for the cell-free synthesis of membrane proteins,
Nature Protocols 10, 1328-1344 (2015).

[Summary] Cell-free gene expression systems are biotechnological tools for the in vitro production of proteins of interest. The addition of membrane vesicles (liposomes) enables the production of membrane proteins, including those in large molecular weight complexes, such as SecYEG translocon or ATP synthase. Here, we describe a protocol for the cell-free synthesis of membrane proteins using the PURE system, and subsequent quantification of products and analyses of membrane localization efficiency, product orientation in the membrane, and complex formation in the membrane. Additionally, measurements of ATP synthase activity are used as an example to demonstrate the functional nature of the cell-free synthesized proteins. This protocol allows the rapid production and the detailed analysis of membrane proteins, and the complete process from template DNA preparation to activity measurement can be accomplished within one day. This protocol can also avoid the risks in membrane protein purification and in reconstruction to lipid membrane, compared to alternative methods using living cells.

2014

Hideaki Matsubayashi, Yutetsu Kuruma, and Takuya Ueda,
Cell-Free Synthesis of SecYEG Translocon as the Fundamental Protein Transport Machinery,
Origins of Life and Evolution of Biospheres 44, 331-334 (2014).

Hideaki Matsubayashi, Yutetsu Kuruma, and Takuya Ueda,
In vitro synthesis of the E. coli Sec Translocon from DNA,
Angewandte Chemie International Edition 53, 7535-7538 (2014).

*Yoshihiro Shimizu, Yutetsu Kuruma, Takashi Kanamori, Takuya Ueda ,
The PURE system for protein production,
Methods in Molecular Biology 1118, 275-284 (2014).




Paper | Proceedings

2015

*Yutetsu Kuruma,
Creation of Simple Biochemical Systems to Study Early Cellular Life,
Origins of Life and Evolution of Biospheres 45, 359-360 (2015).

Pier Luigi Luisi, and Yutetsu Kuruma,
Open Questions on the Origin of Life (OQOL)-Introduction to the Special Issue,
Origins of Life and Evolution of Biospheres 44, 267-268 (2015).




International Conferences

2015

Invited

*Yutetsu Kuruma,
Construction of Self-Reproducing Minimal Cell,
International WorkShop on Challenge to Synthesizing Life (Aug. 25-26, 2015), Hakone, Japan.

Poster

*Yutetsu Kuruma,
Fatty acid synthesis inside giant unilamellar vesicle -Toward the construction of self-reproducing protocell-,
International Symposium on Fluctuation and Structure out of Equilibrium 2015 (SFS2015) (Aug. 20-23, 2015), Kyoto, Japan.

Invited

*Yutetsu Kuruma,
Construction of Self-reproducing Minimal Cell,
Sattelite Workshop “The 1st Workshop on Synthetic Natural Systems”, in ECAL2015 (Jul. 20-24, 2015), York, UK.

Oral (contributed)

*Yutetsu Kuruma,
Reconstruction of cell membrane function,
RNA, Peptides, Vesicles and Exoplanets (Feb. 26-27, 2015), Boston, USA.

Poster

*Hideaki Matsubayashi, Yutetesu Kuruma, Takuya Ueda,
Cell-free synthesis of Sec translocon on liposome membrane,
Biophysical Society 59th Annual Meeting (Feb. 7-11, 2015), Maryland, USA.

*Yutetsu Kuruma,
Construction of artificial cell membrane for the study of the origin and evolution of life,
The 3rd ELSI (Earth-Life Science Institute) International Symposium (Jan. 13-15, 2015), Tokyo, Japan.


2014

Poster

*Yutetsu Kuruma, Hideaki Matsubayashi, and Takuya Ueda,
In Vitro Reconstruction of Functional Membrane,
ALIFE14 (Jul. 30 – Aug. 2, 2014), New York, USA.

Oral (contributed)

*Yutetsu Kuruma,
In vitro reconstruction of functional cell membrane,
Satellite workshop in ALIFE14, What can Synthetic Biology offer to (Embodied) Artificial Intelligence? (Jul. 30, 2014), New York, USA.

Poster

*Hideaki Matsubayashi, Yutetsu Kuruma, Takuya Ueda,
In Vitro Synthesis of Sec Translocon from DNA,
Open Questions on the Origin of Life 2014 (Jul. 12-13, 2014), Nara, Japan.

Oral (contributed)

*Yutetsu Kuruma, Hideaki Matsubayashi, Pasquale Stano, Takuya Ueda, Pier L. Luisi, Daisuke Kiga,
Creation a possible living organism that might exist in an early earth condition,
ORIGINS2014 (Jul. 6-11, 2014), Nara, Japan.




Book

2015

Open Questions on the Origin of Life (OQOL)—Introduction to the Special Issue
Yutetsu Kuruma and P. L. Luisi(編)
Origins of Life and Evolution of Biospheres(2015)


2014

「Artificial Life 14」
“In Vitro Reconstruction of Functional Membrane”
Yutetsu Kuruma, Hideaki Matsubayashi, and Takuya Ueda
The MIT Press(2014), 963-964.

Grant-in-Aid for Scientific Research (KAKENHI) on Innovative Areas, MEXT, Japan
Synergy of Fluctuation and Structure : Quest for Universal Laws in Non-Equilibrium Systems