2012 Fiscal Year Final Research Report
Crystallization of stabilized leukotriene B4 receptor mutant and monoclonal antibody complex.
| Project/Area Number |
23770133
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Structural biochemistry
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
HORI Tetsuya 独立行政法人理化学研究所, 宮野構造生物物理研究室, 専任研究員 (20344054)
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| Project Period (FY) |
2011 – 2012
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| Keywords | ロイコトリエンB_4 受容体 / G-タンパク質共役型受容体 / メタノール資化酵母 / 安定化変異体 / モノクローナル抗体 / 大量発現 / 精製 / 結晶化 |
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| Research Abstract |
For the crystallization of leukotriene B_4 receptor (BLT1), the stabilized mutant was constructed and the anti-BLT1 monoclonal antibody which recognized three-dimensional structure of BLT1 was generated. The rationally designed two point mutations were improved the thermal-stability by 5 ℃ and the expression level of active form BLT1 by 6 times (Bmax=311 pmol/mg). The antibody which bound the none-denatured BLT1 but did not recognize the SDS-denatured BLT1 was generated. The crystallization of BLT1-Fab complex has been promoted.
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