| Project/Area Number |
14370759
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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 |
医薬分子機能学
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| Research Institution | Kumamoto University |
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Principal Investigator |
OTAGIRI Masaki Kumamoto University, Faculty of Medical and Pharmaceutical Sciences, Professor, 大学院・医学薬学研究部, 教授 (80120145)
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| Co-Investigator(Kenkyū-buntansha) |
TANASE Sumio Kumamoto University, Faculty of Medical Sciences, Professor, 医学部, 教授 (20112401)
NOZAKI Syuei The Chemo-Sero-Therapeutic Research Institute, Manager, 菊池研究所, 室長
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2004: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2003: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥6,700,000 (Direct Cost: ¥6,700,000)
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| Keywords | recombinant / albumin preparation / antioxidant activity / long half-life / pharmacokinetics / functional property / structural property / アルブミン製剤 |
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| Research Abstract |
Human serum albumin(HSA) has many essential functions for homeostasis, such as the maintenance of osmotic pressure, drug binding and antioxidant activity. Mutagenesis studies of HSA made it possible to examine the participation of various amino acid residues and domains in the functional properties of HSA, such as binding capacity, antioxidant activity and prolonged half-life.
In this study, firstly, we studied the antioxidant properties of single-residue mutants of human serum albumin. Cysteine residues, especially ^<166>Cys contribute much to this function of albumin. Secondly, Pichia pastoris has been used to express each of the three rHSA domains separately. Recombinant domain I protein exhibited antioxidant activity comparable to that of rHSA. A novel domain exchange HSA with potentially high antioxidant activity was designed where three domain I proteins of HSA were genetically fused. This exchanged HSA had higher antioxidant activity than wild-type HSA. Thirdly, rHSA dimer has been expressed using Pichia pastoris. The structure and binding capacity of rHSA dimer was almost identical to that of native HSA. rHSA dimer was shown to exhibit better intravascular retention and lower vascular permeability properties. Lastly, single-residue mutants R410C efficiently introduced NO further than the wild type. S-NO-R410C had higher antibacterial activity than S-NO-HSA.
The results of this research serve as important fundamental data for further development of new recombinant albumin.
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