著者

Yuki Sudo Rikou Tanaka Toshitatsu Kobayashi Naoki Kamo Toshiyuki Kohno Chojiro Kojima

出版者

The Biophysical Society of Japan

雑誌

BIOPHYSICS (ISSN:13492942)

巻号頁・発行日

vol.7, pp.51-58, 2011 (Released:2011-06-18)

参考文献数

34

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An approach of cell-free synthesis is presented for the functional expression of transmembrane proteins without the need of refolding. The transmembrane region of the pharaonis halobacterial transducer protein, pHtrII, was translated with various large soluble tags added (thioredoxin, glutathione S-transferase, green fluorescent protein and maltose binding protein). In this system, all fusion pHtrII were translated in a soluble fraction, presumably, forming giant micelle-like structures. The detergent n-dodecyl-β-D-maltoside was added for enhancing the solubilization of the hydrophobic region of pHtrII. The activity of the expressed pHtrII, having various tags, was checked using a pull-down assay, using the fact that pHtrII forms a signaling complex with pharaonis phoborhodopsin (ppR) in the membrane, as also in the presence of a detergent. All tagged pHtrII showed a binding activity with ppR. Interestingly, the binding activity with ppR was positively correlated with the molecular weight of the soluble tags. Thus, larger soluble tags lead to higher binding activities. We could show, that our approach is beneficial for the preparation of active membrane proteins, and is also potentially applicable for larger membrane proteins, such as 7-transmembrane proteins.

著者

Kei TAKAKURA Takayuki SAKO Haruo MIYADERA Kenichi YOSHIOKA Yoshiji KARINO Kohichi NAKAYAMA Tsukasa SUGITA Daisuke UEMATSU Kohei OKUTOMO Jun HASEGAWA Toshiyuki KOHNO Eiki HOTTA

出版者

The Japan Society of Plasma Science and Nuclear Fusion Research

雑誌

Plasma and Fusion Research (ISSN:18806821)

巻号頁・発行日

vol.13, pp.2406075, 2018-06-12 (Released:2018-08-22)

参考文献数

9

被引用文献数

5

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Feasibility studies on neutron radiography using an inertial electrostatic confinement (IEC) neutron source were carried out. A Monte Carlo analysis was carried out to evaluate the feasibility of neutron radiography experiment. Imaging tests using a medium-sized IEC neutron source were conducted with the indirect method using a dysprosium (Dy) foil and an imaging plate. Neutron images of objects consisting of six Cd pins and an array of B4C powder contained in a stainless-steel blade were obtained. The numerical and experimental results confirmed that the IEC neutron source can be applied to neutron radiography even with relatively low neutron flux of ∼102 n/s/cm2.