KEIO RESEARCHERS INFORMATION SYSTEM

Career 【 Display / hide 】

Career 【 Display / hide 】

• 2009.04

  -

  2013.03

  名古屋大学, 物質科学国際研究センター

• 2013.04

  -

  2014.03

  名古屋大学, 大学院理学研究科生命理学専攻

• 2014.04

  -

  2017.03

  慶應義塾大学, 理工学部

• 2017.04

  -

  2020.03

  慶應義塾大学, 理工学部, 専任講師（有期）

• 2020.04

  -

  Present

  慶應義塾大学, 理工学部

Academic Background 【 Display / hide 】

Academic Background 【 Display / hide 】

• 2002.04

  -

  2004.03

  Ube National College of Technology, 専攻科物質科学専攻

  Technical College, Completed

• 2004.04

  -

  2006.03

  Hiroshima University, 大学院理学研究科博士課程前期

  Master's course

• 2006.04

  -

  2009.03

  Hiroshima University, 大学院理学研究科博士課程後期

  Doctoral course

Academic Degrees 【 Display / hide 】

Academic Degrees 【 Display / hide 】

• 博士（理学）, Hiroshima University, Coursework, 2009.03

Licenses and Qualifications 【 Display / hide 】

Licenses and Qualifications 【 Display / hide 】

• 放射線取扱主任者第一種, 2015.03

Research Areas 【 Display / hide 】

Research Areas 【 Display / hide 】

• Natural Science / Biogeosciences (金属イオン、金属タンパク質、実験室進化)

• Nanotechnology/Materials / Chemistry and chemical methodology of biomolecules (タンパク質、ナノ粒子、ハイドロゲル)

Research Themes 【 Display / hide 】

Research Themes 【 Display / hide 】

• 生命の元素に対する進化的可塑性の解明, 

  2015.04

  -

  Present

• タンパク質を鋳型とした分子構造の設計と材料への応用, 

  2014.03

  -

  Present

Books 【 Display / hide 】

Books 【 Display / hide 】

• Dual Modification of Artificial Protein Cage

  Kawakami N., Nasu E., Miyamoto K., Methods in Molecular Biology, 2023

  　View Summary

  Chemical modifications of proteins confer new functions on them or modulate their original functions. Although various approaches are developed for modifications, modifications of the two different reactive sites of proteins by different chemicals are still challenging. In this chapter, we show a simple approach for selective modifications of both interior and exterior surfaces of protein nanocages by two different chemicals based on a molecular size filter effect of the surface pores.

  • Access to Document (DOI)

Papers 【 Display / hide 】

Papers 【 Display / hide 】

• misteINK: a protein nanocage-based ink with reversible, stimuli-responsive color shifts

  Yamashita M., Kawakami N., Arai R., Ikeda A., Moriya T., Senda T., Miyamoto K.

  Biomaterials Science 13 ( 23 ) 6652 - 6661 2025.12

  ISSN  20474830

  　View Summary

  Dyes exhibiting polarity-dependent color changes, known as solvatochromism, have great potential for creating sensors, smart materials, and responsive coatings. However, full-range color shifts require a technique to disperse dyes across a wide range of solvent polarities, which remains a persistent challenge. For example, hydrophobic dyes often aggregate in water, preventing effective color shifts. Although surfactants can assist in dye dispersion, they can also prevent solvent molecules from accessing the dye. To address this, we used a 60-mer protein nanocage, TIP60, with a densely pyrene-modified interior surface. The modification did not induce protein denaturation, as monitored by small-angle X-ray scattering, and greatly increased the aqueous solubility of a hydrophobic solvatochromic dye, Nile Red (NR), while preserving its fluorescence. The NR-loaded solution appeared blue, reflecting the polar environment surrounding NR. Cryogenic electron microscopy suggested that the pyrenes interacted with each other to form a binding site for NR. This interaction also contributed to thermostability of TIP60 (65 °C to 86 °C) and stability against sodium dodecyl sulfate, as observed by electrophoresis experiments. When brushed onto plain copy paper, the NR-loaded nanocage appeared bluish-purple and shifted reversibly to purplish red upon heating, returning on cooling—presumably via nanocage dissociation and reassembly. The color change was also sensitive to humidity. We term this material “misteINK”, a protein-based ink with reversible temperature- and humidity-dependent color changes. These findings demonstrate that a single-step interior modification enables the rational design of protein materials for tuning dye photophysics, providing a powerful strategy for designing protein-based functional materials.

  • Access to Document (DOI)

• Design, structures, and applications of an icosahedral artificial protein nanocage, TIP60

  Kawakami N., Miyamoto K., Arai R.

  Chemical Communications 61 ( 53 ) 9584 - 9595 2025.05

  ISSN  13597345

  　View Summary

  Many protein nanocages are polyhedral shapes with icosahedral symmetry owing to the geometric constraints defined by Euler's polyhedral theorem, which suggests that pentagonal faces play a key role in creating uniform structures. This principle has inspired a range of design approaches to create artificial protein nanocages. For example, this review mainly shows design, structures, and applications of TIP60, an artificially designed icosahedral protein nanocage produced through a fusion protein approach incorporating pentameric structures. Protein engineering techniques, such as chemical modifications and mutation strategies for TIP60, have successfully added properties useful for various applications, including drug delivery, nanoreactors, and hydrogels. Recent advances in computational protein design can enable precise structural customization. Therefore, the integration of rational design with targeted modifications is likely to become a powerful approach for developing functionalized proteins.

  • Access to Document (DOI)

• Fusion then fission: splitting and reassembly of an artificial fusion-protein nanocage

  Ohara N., Kawakami N., Arai R., Adachi N., Ikeda A., Senda T., Miyamoto K.

  Chemical Communications 60 ( 34 ) 4605 - 4608 2024.04

  ISSN  13597345

  　View Summary

  A split-protein system is a simple approach to introduce new termini which are useful as modification sites in protein engineering, but has been adapted mainly for monomeric proteins. Here we demonstrate the design of split subunits of the 60-mer artificial fusion-protein nanocage TIP60. The subunit fragments successfully reformed the cage structure in the same manner as prior to splitting. One of the newly introduced terminals at the interior surface can be modified using a tag peptide and green fluorescent protein. Therefore, the termini could serve as a versatile modification site for incorporating a wide variety of functional peptides and proteins.

  • Access to Document (DOI)

• Thermally Reversible Gel-Sol Transition of Hydrogels via Dissociation and Association of an Artificial Protein Nanocage

  Nasu E., Kawakami N., Takamura S., Hotta A., Arai R., Miyamoto K.

  Biomacromolecules 25 ( 4 ) 2358 - 2366 2024.04

  ISSN  15257797

  　View Summary

  Oligomeric protein nanocages often disassemble into their subunits and reassemble by external stimuli. Thus, using these nanocages as cross-linkers for hydrogel network structures is a promising approach to allow hydrogels to undergo stimuli-responsive gel-sol transitions or self-healing. Here, we report hydrogels that show a reversible gel-sol transition resulting from the heat-induced dissociation and reassociation of protein nanocages. The hydrogel contained the 60-mer artificial protein nanocage, TIP60, as a supramolecular cross-linker for polyethylene glycol network structures. The hydrogel showed a gel-to-sol transition upon heating at a temperature above the melting point of TIP60 and immediately returned to a gel state upon cooling to room temperature. During the heating and cooling treatment of the hydrogel, small-angle X-ray scattering analysis suggested the dissociation and reassociation of TIP60. Furthermore, we demonstrated redox-responsive cargo release from TIP60 in the hydrogel. These results showed the potential of TIP60 as a component of multi-stimuli-responsive hydrogels.

  • Access to Document (DOI)

• Column-free purification of an artificial protein nanocage, TIP60

  Nasu E., Kawakami N., Ohara N., Hayashi K., Miyamoto K.

  Protein Expression and Purification (Protein Expression and Purification)  205 2023.05

  ISSN  10465928

  　View Summary

  Protein nanocages, which have inner cavities and surface pores, are attractive materials for various applications, such as in catalysts and medicine. Recently, we produced an artificial protein nanocage, TIP60, and demonstrated its potential as a stimuli-responsive nanocarrier. In the present study, we report a simple purification method for TIP60 that can replace time-consuming and costly affinity chromatography purification. TIP60, which has an anionic surface charge, aggregated at mildly acidic pH and redissolved at neutral pH, maintaining its cage structure. This pH-responsive reversible precipitation allowed us to purify TIP60 from soluble fractions of the E. coli cell lysate by controlling the pH. Compared with conventional Ni-NTA column purification, the pH-responsive precipitation method provided purified TIP60 with similar purity (∼80%) and higher yield. This precipitation purification method should facilitate the large-scale investigation and practical use of TIP60 nanocages.

  • Access to Document (DOI)

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Papers, etc., Registered in KOARA 【 Display / hide 】

Papers, etc., Registered in KOARA 【 Display / hide 】

• 気楽にやりませんか?

  Kawakami, Norifumi

  新版 窮理図解 (慶應義塾大学理工学部)   ( 37 )  2023.08

• Why not start small?

  Kawakami, Norifumi

  New Kyurizukai (Faculty of Science and Technology, Keio University)   ( 37 )  2023.08

• Evolution experiment of Escherichia coli in the presence of abiotic metal ions

  Kawakami, Norifumi

  科学研究費補助金研究成果報告書 2023

• Use of the artificial protein nanocage as nanoscale phase separator

  Kawakami, Norifumi

  科学研究費補助金研究成果報告書 2021

• Chemical modification of hollow artificial protein nano particle for incorporation of small molecules

  Kawakami, Norifumi

  学事振興資金研究成果実績報告書 (慶應義塾大学)  2018

Research Projects of Competitive Funds, etc. 【 Display / hide 】

Research Projects of Competitive Funds, etc. 【 Display / hide 】

• 実験室進化で生じた大腸菌変異株の鉄輸送体に着目した進化プロセスの検証

  2022.04

  -

  2024.03

  Research grant, Principal investigator

• 大腸菌実験室進化を通じた生物の元素利用に対する進化的可塑性の検討

  2021.07

  -

  2024.03

  MEXT,JSPS, Grant-in-Aid for Scientific Research, Grant-in-Aid for Challenging Research (Exploratory), Principal investigator

• 人工球状タンパク質超分子を用いたナノスケール相分離の実現とその応用

  2018.04

  -

  2021.03

  MEXT,JSPS, Grant-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Principal investigator

Awards 【 Display / hide 】

Awards 【 Display / hide 】

• 第8回新化学技術研究奨励賞 新化学技術推進協会

  2019.06

  Type of Award： Award from publisher, newspaper, foundation, etc.

• 日本蛋白質科学会若手奨励賞優秀賞

  2017.06

Courses Taught 【 Display / hide 】

Courses Taught 【 Display / hide 】

• INTRODUCTION TO BIOLOGY

  2026

• OFF-CAMPUS STUDY PROGRAM 2

  2026

• BASIC LABORATORY COURSE IN BIOSCIENCES

  2026

• BIOTRANSFORMATION (BIOLOGICAL CHEMISTRY 4)

  2026

• OFF-CAMPUS STUDY PROGRAM 1

  2026

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Courses Previously Taught 【 Display / hide 】

Courses Previously Taught 【 Display / hide 】

• 生体反応論第2

  Keio University

  2017.04

  -

  Present

  , 

  Spring Semester