Development of a novel system for solubilization and preparation of proteins based on the aggregation suppression effects of arginine on proteins
| Project/Area Number |
17360393
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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 |
Biofunction/Bioprocess
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TSUMOTO Kouhei The University of Tokyo, Graduate School of Frontier Sciences, Associate Professor (90271866)
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| Co-Investigator(Kenkyū-buntansha) |
UMETSU Mitsuo Tohoku University, Graduate School of Engineering, Associate Professor (70333846)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,430,000 (Direct Cost: ¥15,500,000、Indirect Cost: ¥930,000)
Fiscal Year 2007: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2006: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2005: ¥8,300,000 (Direct Cost: ¥8,300,000)
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| Keywords | PROTEIN / BIOTECHNOLOGY / BIOMOLECULE / BIO-ENGINEERING / 溶媒・添加剤 / アルギニン / 可溶化 / クロマトグラフィー / 凝集会合形成 / フィロールディング / 製剤 / プロテインマニピュレーション / リフォールディング |
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| Research Abstract |
To establish the novel system for solublization and preparation of proteins using amino acids and their derivatives, we have focused on the characteristics of arginine and it derivatives; mechanism on protein aggregation suppression of arignine and its derivatives and its application to several bioprocesses for proteins have been addressed from the following viewpoints.
1. Elucidation of aggregation suppression mechanism on proteins of L-arginine and its derivatives Solubility measurements of L arginine and its derivatives with various amino acids, and peptide bonds have indicated the identical affinities for main chain and side chains to those of guanidium ion. Several derivatives could improve the specific characteristics of L-arginine, e.g. efficiency of solubilization of proteins. 2) Crystal structures of two proteins under existence of L-arginine: crystal structures of two proteins have indicated that L-arginine made critical contribution to the stabilization of hydrated proteins,
… More
and also suggested that the mechanism on the proteins is dependent upon the tertiary structures of target proteins.
2. Solubilization of proteins using L-arginine and its derivatives E-coli expressed signal-transduction-related proteins, and membrane proteins could be solubilized from inclusion bodies by using L-arginine and its derivatives. Thermodynamics analyses have revealed the complete recovery of the functions and structures of the solubilized proteins. We could successfully establish the method for solubilization of proteins using L-arginine and its derivatives.
3. Application of arigine to solvent additives of chromatographies: it could be shown that arginine could be utilized for an effective solvent additive of various chromatographies, including GPC, HIC, IEX, and several affinity chromatographies.
From these results, our achievements could be summarized as follows; the mechanism of arginine on protein structure, effective utilization of arginine for solubilzation and purification of proteins. Less
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Report
(4 results)
Research Products
(227 results)
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[Journal Article] Critical contribution of aromatic rings to specific recognition of polyether rings : The case of ciguatoxin CTX3C-ABC and its specific antibody 1C492008
Author(s)
Tsumoto K^*, Yokota A, Tanaka Y, Ui M, Tsumuraya T, Fujii I, Kumagai I, Nagumo Y, Oguri H, Inoue M, Hirama M.
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A helical string of alternately connected three-helix bundles for the cell wall-associated adhesion protein Ebb from Staphylococcus aureus2008
Author(s)
Tanaka Y, Sakamoto S, Kuroda M, Goda S, Gao YG, Tsumoto K, Hiragi Y, Yao M, Watanabe N, Ohta T, Tanaka I.
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Journal Title
Structure 16(3)
Pages: 488-496
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Entropically driven MHC class I recognition by human inhibitory receptor leukocyte Ig-like receptor B1 (LILRBI/ILT2/CD85j)2006
Author(s)
Shiroishi M, Kuroki K, Tsumoto K, Yokota A, Sasaki T, Amano K, Shimojima T, Shirakihara Y, Rasubala L, van der Merwe PA, Kumagai I, Kohda D, Maenaka K.
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Journal Title
J. Mol. Biol. 355
Pages: 237-248
Description
「研究成果報告書概要(欧文)」より
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[Presentation] 癌免疫療法の現状と今後2007
Author(s)
津本
Organizer
日本癌学会シンポジウム
Place of Presentation
パシフィコ横浜
Year and Date
2007-10-03
Description
「研究成果報告書概要(和文)」より
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[Book] たんぱく質結晶の新展開2008
Author(s)
津本、田中
Total Pages
15
Publisher
シーエムシー出版
Description
「研究成果報告書概要(和文)」より
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[Book] ナノバイオ辞典2006
Author(s)
津本
Total Pages
2
Publisher
テクノシステム
Description
「研究成果報告書概要(和文)」より
Related Report
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[Book] バイオプロダクション2006
Author(s)
津本
Total Pages
20
Publisher
コロナ社
Description
「研究成果報告書概要(和文)」より
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