A03 NISHIZAKA, Takayuki |Proposed Research Projects (2016-2017)

Paper | Original Paper


Masaki Mizutani, Isil Tulum, Yoshiaki Kinosita, Takayuki Nishizaka, Makoto Miyata,
Detailed Analyses of Stall Force Generation in Mycoplasma mobile Gliding,
Biophysical Journal 114, 1411-1419 (2018).

Yoshiaki Kinosita, Yoshitomo Kikuchi, Nagisa Mikami, Daisuke Nakane, Takayuki Nishizaka,
Unforeseen swimming and gliding mode of an insect gut symbiont, Burkholderia sp. RPE64, with wrapping of the flagella around its cell body,
The ISME Journal 12, 838-848 (2018).


Daisuke Nakane and Takayuki Nishizaka*,
Asymmetric distribution of type IV pili triggered by directional light in unicellular cyanobacteria,
Proceedings of the National Academy of Sciences 114, 6593-6598 (2017).

[Summary] The type IV pili (T4P) system is a supermolecular machine observed in prokaryotes. Cells repeat the cycle of T4P extension, surface attachment, and retraction to drive twitching motility. Although the properties of T4P as a motor have been scrutinized with biophysics techniques, the mechanism of regulation remains unclear. Here we provided the framework of the T4P dynamics at the single-cell level in Synechocystis sp. PCC6803, which can recognize light direction. We demonstrated that the dynamics was detected by fluorescent beads under an optical microscope and controlled by blue light that induces negative phototaxis; extension and retraction of T4P was activated at the forward side of lateral illumination to move away from the light source. Additionally, we directly visualized each pilus by fluorescent labeling, allowing us to quantify their asymmetric distribution. Finally, quantitative analyses of cell tracking indicated that T4P was generated uniformly within 0.2 min after blue-light exposure, and within the next 1 min the activation became asymmetric along the light axis to achieve directional cell motility; this process was mediated by the photo-sensing protein, PixD. This sequential process provides clues toward a general regulation mechanism of T4P system, which might be essentially common between archaella and other secretion apparatuses.

Shoko Fujimura, Yuko Ito, Mitsunori Ikeguchi, Kengo Adachi, Junichiro Yajima, Takayuki Nishizaka,
Dissection of the angle of single fluorophore attached to the nucleotide in corkscrewing microtubules,
Biochemical and Biophysical Research Communications 485, 614-620 (2017).


Yoshiaki Kinosita, *Nariya Uchida, Daisuke Nakane, and *Takayuki Nishizaka,
Direct observation of rotation and steps of the archaellum in the swimming halophilic archaeon Halobacterium salinarum,
Nature Microbiology 1, 16148/1-9 (2016).

Akihiro Tanaka, Daisuke Nakane, Masaki Mizutani, Takayuki Nishizaka, *Makoto Miyata,
Directed binding of gliding bacterium, Mycoplasma mobile, shown by detachment force and bond lifetime,
mBio 7, e00455-16 (2016).

*Mitsuhiro Sugawa, Kei-ichi Okazaki, Masaru Kobayashi, Takashi Matsui, Gerhard Hummer, Tomoko Masaike, and *Takayuki Nishizaka,
F1-ATPase conformational cycle from simultaneous single-molecule FRET and rotation measurements,
Proceedings of the National Academy of Sciences of the United States of America 113, E2916-2924 (2016).

International Conferences



*Takayuki Nishizaka, Yoshiaki Kinosita, Takanobu. A. Katoh, Daisuke Nakane,
Structural and functional dynamics of nanoscale molecular motors,
The 31st International Congress on High-Speed Imaging and Photonics (Nov. 7-10, 2016), Osaka, Japan.

*Takayuki Nishizaka,
Visualization of dynamics of motors in archaella and type IV pili system,
Current and Future Perspectives in Active Matter (Oct. 28-29, 2016), Tokyo, Japan.


Kuzuu, S., Daisuke Nakane, *Y. Fukumori, A. Taoka, Takayuki Nishizaka,
Polarity of a bacterial nano-compass reversed by magnetic tweezers,
Magnetotactic Bacteria 5th International Meeting (Sep. 11-15, 2016), Provence, France.

*Kawamura, S., Y. Takaoka, A. Taoka, Daisuke Nakane, Takayuki Nishizaka, Y. Fukumori,
Visualization of flagellar rotation during magnetotactic motility using Magnetospirillum magneticum AMB-1,
Magnetotactic Bacteria 5th International Meeting (Sep. 11-15, 2016), Provence, France.

*Masaki Mizutani, Isil Tulum, Yoshiaki Kinosita, Takayuki Nishizaka, Makoto Miyata,
Force generated by single unit in Mycoplasma mobile gliding,
21st Congress of the International Organization for Mycoplasmology (Jul. 3-7, 2016), Brisbane, Australia.

*Masaki Mizutani, Isil Tulum, Yoshiaki Kinosita, Takayuki Nishizaka, Makoto Miyata,
Force generation based on novel energy conversion system in Mycoplasma mobile measured by laser trap,
7th OCARINA International Symposium in conjunction with Symposium in honor of Distinguished Professor Michael Nobel (Mar. 17-18, 2016), Osaka, Japan.

*Nagisa Mikami, Yuko Ito, Mitsuhiro Sugawa, Mitsunori Ikeguchi, Takayuki Nishizaka,
Conformational change of ab-interface in F1-ATPase detected by anisotropy of fluorescent nucleotide attached to catalytic sit,
Gordon Research Conference (Mar. 3-9, 2016), Hong Kong, China.

Nagisa Mikami, Yuko Ito, K. Adachi, Mitsunori Ikeguchi, *Takayuki Nishizaka,
Conformational Change in γ120 and γ240 of F1-ATPase from the Aspect of Difference of Nucleotides by Advanced TIRF Microscope,
Biophyscial Society 60th Annual Meeting (Feb. 27- Mar. 2, 2016), Los Angeles, USA.

Oral (contributed)

Yoshiaki Kinosita, Daisuke Nakane, Nariya Uchida, Makoto Miyata, *Takayuki Nishizaka,
Unitary Steps of Supermolecular Motility Machineries in Gliding Bacteria and Swimming Arcaea,
Biophyscial Society 60th Annual Meeting (Feb. 27- Mar. 2, 2016), Los Angeles, USA.


Takanobu A. Katoh, Koji Ikegami, Nariya Uchida, Toshihito Iwase, Tomoko Masaike, Mitsutoshi Setou, *Takayuki Nishizaka,
Separating Ciliary Beating Force into Dynein-Driven Force and Axoneme-Intrinsic Bending Force By 3-D Tracking Microscopy,
Biophyscial Society 60th Annual Meeting (Feb. 27- Mar. 2, 2016), Los Angeles, USA.



「Handbook of Photonics for Biomedical Engineering」Aaron Ho-Pui HoDonghyun KimMichael G. Somekh (ed.)
“3-D Single Particle Tracking Using Dual Images Divided by Prism: Method and Application to Optical Trapping” Takanobu A. Katoh, Shoko Fujimura, Takayuki Nishizaka
Springer(2017), 755-766, ISBN : 978-94-007-5051-7

[summary] We here describe the three-dimensional optical tracking method, which is realized with a simple optical component, a quadrangular wedge prism. Additional two lenses located between a conventional optical microscope and a camera enable to track single particles in 3-D. Because of the simplicity of its rationale and construction, any laboratory equipped with 2-D tracking method, under either fluorescence, phase-contrast, bright-field or dark-field illumination, can adopt our method with the same analysis procedure and thus the same precision. Applications to a molecular motor, kinesin-microtubule system, and optical trapping, are also demonstrated, verifying the advantage of our approach to assess the movement of tiny objects, with the size ranging from ten nanometers to a few microns, in an aqueous solution.
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