| Project/Area Number |
61480452
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
結晶学
|
|---|
| Research Institution | University oif Tokyo |
|---|
Principal Investigator |
IITAKA Yoichi Professor at the University of Tokyo, 薬学部, 教授 (90012591)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KASAI Hisataka Associate Professor at Tokyo Metroopolitan University, 理学部, 助教授 (80087163)
NAKAMURA Kazuo Instructur at the University of Tokyo, 薬学部, 助手 (00012675)
MITSUI Yukio Instructor at the University of tokyo, 薬学部, 助手 (40012637)
|
|---|
| Project Period (FY) |
1986 – 1987
|
| Project Status |
Completed (Fiscal Year 1987)
|
| Budget Amount *help |
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1987: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1986: ¥2,900,000 (Direct Cost: ¥2,900,000)
|
|---|
| Keywords | Calmodulin / Crystallography / X-ray Diffraction Drug Action / Melittin / メリチン / 生理活性ペプチド / X線解析 / 結晶学的研究 / 向精神薬 / オリゴペプチド |
|---|
| Research Abstract |
Abbreviation : CaM = Calmodulin
(1) Crystallization of various complexes : a)Reproducible conditions for growing Ca^<2+>-CaM-chlorpromazine ternary complex was established. b) Also, Ca^<2+>-CaM-melittin ternary complex was crystallized and crystallogrpahically characterized. c)For Ca^<2+>-CaM-mastoparan ternary complex, only spherulites were obtained.
(2) Attempt at X-ray structure analysis of Ca^<2+>-CaM-Chlorpromazine complex : a) At the outset of this project, we predicted that the unusually long c-axis (178 A) can be a serious obstacle for data collection. The actual data collection, in fact, showed that only the data up to 4 A resolution can be usable for structure analysis. Another attempt using the synchrotoron radiation X-ray source, which has much narrower beam divergence, failed due to large temperature rise inside the experimentation hatch. Data collection using some kind of two-dimensional detector(s) are being planned. b) Regarding the heavy-atom derivatives for solving the phase problem, the K_2PtCl_4 and Pb(CH_3COO)_2 derivatives were proved to be promising.
(3) Separation of the complexes through the use of ion exchange HPLC : ONLY the Ca^<2+>-CaM-melittin complex was detcted on the HPLC patterm using DEAE-5PW column, indicating that significant binding forces were not existent in the complexes with mastoparan.
(4) Some biochemical studies on melitttin, mastoparan. : a) Melitttin as well as mastoparan binds to CaM-Sepharose column, the latter peptide being slightly weaker ligand. b) Hemolytic activity observed for melittin was not observed for mastoparan.
|
