2004 Fiscal Year Final Research Report Summary
Design of bio-nanosystem by dynamic control of nanoorganization
| Project/Area Number |
15300158
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
AKIYOSHI Kazunari Tokyo Medical and Dental University, Institute of Biomaterials and Bioengineering, Professor, 生体材料工学研究所, 教授 (90201285)
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| Co-Investigator(Kenkyū-buntansha) |
IWASAKI Yasuhiko Tokyo Medical and Dental University, Institute of Biomaterials and Bioengineering, Associate Professor, 生体材料工学研究所, 助教授 (90280990)
WATANABE Akihiko Tokyo Medical and Dental University, Institute of Biomaterials and Bioengineering, Assistant Professor, 生体材料工学研究所, 助手 (30126263)
MORIMOTO Nabuyuki Tokyo Medical and Dental University, Institute of Biomaterials and Bioengineering, Assistant Professor, 生体材料工学研究所, 助手 (00313263)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Nanogel / Associated polymer / Self-assembly / Hydrophobized polysaccharides / Stimuli-responsive / Artificial molecular chaperone / Protein folding |
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| Research Abstract |
The research could create novel bio-nanosystem with the function of artificial molecular chaperone for biotechnology and medicine by dynamic control of nanogel particles. In particular, 1.Nanogels, composed of cholesterol bearing pullulan (CUP), -cyclodextrin (CD) systems have been used in order to assist with protein refolding as a novel molecular chaperone-inspired system. The CHP nanogels selectively trapped non-native proteins and CD acted as an effector molecule to control the binding ability of the host to proteins. It indicated the possibility to design various functional nanogels and to apply for refolding chromatography and batchwise renaturation. 2.Dynamic CHP-CD nanogels were characterized by SEC and SEC-MALS methods. The nanogels prevented the thermal aggregation of protein by selective trapping of the heat-denatured protein. After the complex between the CHP-CD nanogels and protein was cooled, the enzyme activity of protein spontaneously recovered upon release from the complex. The dynamic nanogels self-regulated an association of heat denatured protein and dissociation of native protein depending on the concentration of CD. The thermal stability of CAB was improved by thermoresponsive controlled association between the proteins and the artificial molecular chaperone. This is a novel example of a thermoresponsive controlled association between a protein and an artificial chaperone. CHP-CD nanogel systems are useful as protein stabilized reagents. 3.Novel photoresponsive nanogels were prepared by the self-assembly of spiropyrane-bearing pullulan (SpP). The solution properties of the nanogels could be controlled by photostimulation via isomerization between hydrophobic spiropyrane and hydrophilic merocyanine. The molecular chaperone-like activity of the nanogels in protein refolding was investigated. The activity of citrate synthase significantly increased when the amphiphilicity of SpP nanogels was switched by photostimulation.
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