Elucidation of catalytic mechanisms as well as subunit-assembly of nitrile hydratase family enzymes by using advanced structural studies
| Project/Area Number |
24350082
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Partial Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Research Field |
Chemistry related to living body
|
|---|
| Research Institution | Akita University (2014)
Tokyo University of Agriculture and Technology (2012-2013) |
|---|
Principal Investigator |
ODAKA Masafumi 秋田大学, 工学(系)研究科(研究院), 教授 (20224248)
|
|---|
| Co-Investigator(Renkei-kenkyūsha) |
KUROKI Ryota 独立行政法人日本原子力研究開発機構, 量子ビーム応用研究部門, ユニット長 (30391246)
YOHDA Masafumi 東京農工大学, 大学院工学研究院, 教授 (50250105)
|
|---|
| Project Period (FY) |
2012-04-01 – 2015-03-31
|
| Project Status |
Completed (Fiscal Year 2014)
|
| Budget Amount *help |
¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2014: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2012: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
|
|---|
| Keywords | 時間分割結晶構造解析 / 翻訳後修飾 / システインスルフェン酸 / 酵素反応 / 触媒機構 / 反応中間体 / 水和反応 / 結晶構造 / 酵素触媒機構 / 金属酵素 / 構造生物学 / 中性子構造解析 / 酵素反応機構 / 時分割結晶構造解析 |
|---|
| Outline of Final Research Achievements |
Catalytic reaction of a Fe-type nitrile hydratase betaR56K mutant was studied by time-resolved crystallography. The results showed that the metal-coordinated substrate is nucleophilically attacked by the O(SO-) atom of alphaCys114-SO-;, followed by nucleophilic attack of the S(SO-) atom by a betaArg56-activated water molecule to release the product amide and regenerate alphaCys114-SO-;. Also, kinetic as well as crystallographic studies on the mutant SCNases revealed that both NHase and SCNase share the catalytic mechanism and that the size and shape of their substrate binding pockets are predominantly control the substrate selectivity.
|
Report
(4 results)
Research Products
(24 results)
-
[Journal Article] Carbonyl sulfide hydrolase from Thiobacillus thioparus strain THI115 is one of the β-carbonic anhydrase family enzymes2013
Author(s)
Ogawa T, Noguchi K, Saito M, Nagahata Y, Kato H, Ohtaki A, Nakayama H, Dohmae N, Matsushita Y, Odaka M, Yohda M, Nyunoya H, Katayama Y
-
Journal Title
J. Am. Chem. Soc.
Volume: 135
Issue: 10
Pages: 3818-3825
DOI
Related Report
Peer Reviewed
-
[Journal Article] Two arginine residues in the substrate pocket predominantly control the substrate selectivity of thiocyanate hydrolase2013
Author(s)
Yasuaki Yamanaka, Takatoshi Arakawa, Toshinori Watanabe, Satoshi Namima, Masa Sato, Shota Hori, Akashi Ohtaki, Keiichi Noguchi, Yoko Katayama, Masafumi Yohda, Masafumi Odaka
-
Journal Title
Journal of Bioscience and Bioengineering
Volume: Vol. 116、Issue 1
Issue: 1
Pages: 22-27
DOI
NAID
Related Report
Peer Reviewed
-
-
[Presentation] Reaction mechanism of nitrile hydratase proposed by the high resolution transition state structure of the enzyme-substrate complex2014
Author(s)
Yasuaki Yamanaka, Yuki Kato, Koichi Hashimoto, Keisuke Iida, Kazuo Nagasawa, Hiroshi Nakayama, Naoshi Dohmae, Keiichi Noguchi, Takumi Noguchi, Masafumi Yohda, Masafumi Odaka
Organizer
7th Asian Biological Inorganic Chemistry Conference
Place of Presentation
Surfers Paradise, Gold Coast
Year and Date
2014-11-30 – 2014-12-05
Related Report
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
