Crystallization of stabilized leukotriene B4 receptor mutant and monoclonal antibody complex.
Project/Area Number |
23770133
|
Research Category |
Grant-in-Aid for Young Scientists (B)
|
Allocation Type | Multi-year Fund |
Research Field |
Structural biochemistry
|
Research Institution | The Institute of Physical and Chemical Research |
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)
|
Project Period (FY) |
2011 – 2012
|
Project Status |
Completed (Fiscal Year 2012)
|
Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2011: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
|
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 / ロイコトリエンB4受容体 / 安定化 / モノクロ抗体作成 / GPCR / 熱安定化 / 大量調製 |
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.
|
Report
(3 results)
Research Products
(4 results)