| Project/Area Number |
15570133
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Biophysics
|
|---|
| Research Institution | Chiba University |
|---|
Principal Investigator |
ITO Kohji Chiba University, Research associate, 理学部, 助手 (50302526)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Keiichi Chiba University, Department of Biology, Professor, 理学部, 教授 (70053361)
UYEDA Taro Q.P. AIST, Gene Function Reserch Laboratory, director, ジーンファンクション研究センター, 主任研究員副センター長 (90356551)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
|
|---|
| Keywords | mosin / motor / Characean / Cytoplasmic streaming / ATPas activity / in vitro motility assay / actin / in vitro motility assay / 分子モーター |
|---|
| Research Abstract |
The mechanism and structural features that are responsible for the fast motility of Chara corallina myosin(CCM) have not been elucidated, so far. The low yields of native CCM that can be purified to homogeneity were the major reason for this. Here, we describe the expression of recombinant CCM motor domains, which support the fast movement of actin filaments in an in vitro motility assay. A CCM motor domain without light chain binding site moved actin filaments at a velocity of 8.8 μm/s at 30℃ and a CCM motor domain with an artificial lever arm consisting of two alpha-actinin repeats moved actin filaments at 16.2 μm/s. Both constructs displayed high actin-activated ATPase activities (〜500 Pi/s/head), which is indicative of a very fast hydrolysis step. Our results provide an excellent system to dissect the specific structural and functional features that distinguish the myosin responsible for fast cytoplasmic streaming.
|
