| Project/Area Number |
05454629
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Functional biochemistry
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
INOUE Akio Osaka University Grad.school of Science Assistant professor, 理学部, 講師 (80107060)
|
|---|
| Project Period (FY) |
1993 – 1995
|
| Project Status |
Completed (Fiscal Year 1995)
|
| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥5,000,000 (Direct Cost: ¥5,000,000)
|
|---|
| Keywords | Muscle contraction / Myosin / Myosin head / ATPase / 筋肉 / S-1 / 抗体 / 蛋白質構造 / 生体運動 / 分子モーター / プペチド抗体 / 骨格筋 |
|---|
| Research Abstract |
We prepared antibodies specific to heads A and B of myosin using the difference in the amino-acid sequences of two heads of myosin. We studied whether one myosin molecula has A-B heterodimer structure. It has shown that there exists two defferent heads, A and B.We fixed antibodies thus obtained to protein-A which binds to insoluble materials, and examined whether myosin or S-1 (myosin head) can bind with these antibodies. We found that almost all the myosin added was absorbed by either anti-A or anti-B antibody. Therefore, we concluded that most of the myosin molecule has A-B heterodimer structure. The difference in the chemical structure between heads A and B of the myosin molecule has not yet been elucidated, except the amino-acid sequence around 86th reactive lysine residue. Then, we studied the tryptic digestion of s-1 using antibodies specific to two heads. When S-1 was digested by trypsin myosin heavy chain (100 kDa) was digested into three fragments with molecular masses of 25,50, and 21 kDa. Only 25 kDa peptide which contain the reactive lysine residue. was stained by the antibodies. The 25 kDa peptide was formed via 27 kDa peptide, which is the complex of 25 KDa peptidew and 2 kDa junction peptide. The 27 kDa peptide was stained only by anti-B antibodies. This result suggested that the structure around 25 k-50 k junction differs between heads A and B.Since the ATPase active site was located between the domains of 25 kDa and 50 kDa, the structure of ATPase active site may differs between heads A and B.
|
