| Project/Area Number |
21300110
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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 |
Bioinformatics/Life informatics
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
KURODA Yutaka 東京農工大学, 大学院・工学研究院, 准教授 (10312240)
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| Co-Investigator(Kenkyū-buntansha) |
TOH Hiroyuki 産業技術総合研究所, 生命情報工学研究センター, 副研究センター長 (70192656)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000)
Fiscal Year 2011: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2010: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
Fiscal Year 2009: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
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| Keywords | タンパク質凝集 / 会合自由エネルギー / 溶解度測定 / 溶解度向上タグ / 熱力学 / 単純化タンパク質 / ホスト・ゲスト法 / 蛋白質凝集 / 回帰解析 / SEPタグ |
|---|
| Research Abstract |
Protein solubility is usually characterized in terms of a hydrophobicity scale, which refers to the free energy of transfer of a molecule from an aqueous to a nonpolar solution and is not a "solubility propensity scale" per se. Here, we measured the effects of short poly-amino-acid tags(guests) on the solubility of a host protein, a simplified bovine pancreatic trypsin inhibitor(BPTI), to which they were fused at the C-terminus. We analyzed 10 amino acid types, representing the full range of biophysical properties(acidic, basic, polar, and hydrophobic). As anticipated, positively charged residues significantly increased the solubility of the model protein, at both pH 4. 7 and 8. 7, whereas very hydrophobic poly-Ile markedly reduced the solubility of BPTI. Furthermore, to ensure that the measured solubility values were context independent and could provide a genuine "solubility propensity scale", we confirmed that the tags remained independent of the structure, thermal stability, and biochemical activity of the host protein. These observations suggest that this approach is valuable for measuring the solubility propensities of amino acids, which could eventually allow the calculation of a polypeptide's relative solubility from its amino acid sequence.
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