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Featured researches published by Taketoshi Kambara.


Experimental Cell Research | 2008

Intracellular imaging of targeted proteins labeled with quantum dots

Jungwoo Yoo; Taketoshi Kambara; Kohsuke Gonda; Hideo Higuchi

We developed a new method for imaging the movement of targeted proteins in living cancer cells with photostable and bright quantum dots (QDs). QDs were conjugated with various molecules and proteins, such as phalloidin, anti-tubulin antibody and kinesin. These bioconjugated QDs were mixed with a transfection reagent and successfully internalized into living cells. The movements of individual QDs were tracked for long periods of time. Phalloidin conjugated QDs bound to actin filaments and showed almost no movement. In contrast, anti-tubulin antibody conjugated QDs bound to microtubules and revealed dynamic movement of microtubules. Kinesin showed an interesting behavior whereby kinesin came to be almost paused briefly for a few seconds and then moved once again. This is in direct contrast to the smoothly continuous movement of kinesin in an in vitro assay. The maximum velocity of kinesin in cells was faster than that in the in vitro assay. These results suggest that intracellular movement of kinesin is different from that in the in vitro assay. This newly described method will be a powerful tool for investigating the functions of proteins in living cells.


Journal of Cell Biology | 2018

Kinesin-binding–triggered conformation switching of microtubules contributes to polarized transport

Tomohiro Shima; Manatsu Morikawa; Junichi Kaneshiro; Taketoshi Kambara; Shinji Kamimura; Toshiki Yagi; Hiroyuki Iwamoto; Sotaro Uemura; Hideki Shigematsu; Mikako Shirouzu; Taro Ichimura; Tomonobu M. Watanabe; Ryo Nitta; Yasushi Okada; Nobutaka Hirokawa

Kinesin-1, the founding member of the kinesin superfamily of proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here, we report that kinesin-1 binding changes the microtubule lattice and promotes further kinesin-1 binding. This high-affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low-affinity state by washing out the binding kinesin-1 or by the binding of non-hydrolyzable ATP analogue AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy, and second-harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin-1 to GDP microtubules changes the conformation of the GDP microtubule to a conformation resembling the GTP microtubule.


Journal of Cell Biology | 2018

Conformational switching of microtubule and cooperative binding of kinesin-1 as a base for polarized transport

Tomohiro Shima; Manatsu Morikawa; Junichi Kaneshiro; Taketoshi Kambara; Shinji Kamimura; Toshiki Yagi; Hiroyuki Iwamoto; Sotaro Uemura; Hideki Shigematsu; Mikako Shirouzu; Taro Ichimura; Tomonobu M. Watanabe; Ryo Nitta; Yasushi Okada; Nobutaka Hirokawa


The Japanese Biochemical Society/The Molecular Biology Society of Japan | 2017

Single-molecular mechanism of cytoplasmic dynein head

Yoshimi Kinoshita; Kaori Nishikawa; Taketoshi Kambara; Motoshi Kaya; Hideo Higuchi


Biophysical Journal | 2017

The Power Stroke Distance of Human Cytoplasmic Dynein

Yoshimi Kinoshita; Taketoshi Kambara; Kaori Nishikawa; Motoshi Kaya; Hideo Higuchi


生物物理 | 2013

3P159 細胞質ダイニンの生物物理学的・生化学的解析(11.分子モーター,ポスター,日本生物物理学会年会第51回(2013年度))

Taketoshi Kambara; Yoshimi Kinoshita; Takayuki Nakayama; Hideo Higuchi


生物物理 | 2013

3P158 ヒト細胞質ダイニンのパワーストローク測定(11.分子モーター,ポスター,日本生物物理学会年会第51回(2013年度))

Yoshimi Kinoshita; Taketoshi Kambara; Satoshi Ikeda; Hideo Higuchi


Seibutsu Butsuri | 2013

3P159 Biophysical and Biochemical characterization of human cytoplasmic dynein(11. Molecular motor,Poster)

Taketoshi Kambara; Yoshimi Kinoshita; Takayuki Nakayama; Hideo Higuchi


Seibutsu Butsuri | 2013

3P158 Power Stroke Measurement of Human Cytoplasmic Dynein(11. Molecular motor,Poster)

Yoshimi Kinoshita; Taketoshi Kambara; Satoshi Ikeda; Hideo Higuchi


生物物理 | 2012

2A1522 外部負荷存在下でのヒト細胞質ダイニンのメカノケミカルサイクル(分子モーターII:ダイニン,口頭発表,日本生物物理学会第50回年会(2012年度))

Taketoshi Kambara; Yoshiaki Tani; Motoshi Kaya; Tomohiro Shima; Hideo Higuchi

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