| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥570,000)
Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
In this study, we performed a 100-ns molecular dynamics simulation of the 3α-hydroxysteroid dehydrogenase from Pseudomonas sp. B-0831 (Ps3αHSD) to elucidate the flexibility and mobility of the substrate-binding loop. During the simulation, the structure of the holo-subunit was kept unchanged because of NADH interaction with the substrate-binding loop. On the other hand, the substrate-binding loop in the apo-subunit opened and fluctuated largely. By careful observation of the simulation, we designed the next mutagenesis study and found that T188A mutant lost the ability of loop-helix transition and negative cooperativity for NADH binding. These results suggest that high mobility of the substrate-binding loop could produce Ps3αHSD with characteristic activities such as negative cooperativity for NADH binding.
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