Analysis of 3-dimentional structure β-adrenoceptor ligands and receptor complex by NMR
| Project/Area Number |
15590239
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General pharmacology
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| Research Institution | Niigata University of Pharmacy and Applied Life Sciences |
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Principal Investigator |
NAGATOMO Takafumi Niigata University of Pharmacy and Applied Life Sciences, Dept of Pharmacology, Professor, 薬学部, 教授 (60121240)
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| Co-Investigator(Kenkyū-buntansha) |
OHNUKI Toshio Niigata University of Pharmacy and Applied Life Sciences, Dept of Pharmacology, Assistant Professor, 薬学部, 助手 (60288230)
KANEKO Kimiyoshi Niigata University of Pharmacy and Applied Life Sciences, Dept of Pharmacology, Associate Professor, 薬学部, 助教授 (90133462)
片山 肇 新潟薬科大学, 薬化学教室, 教授 (70113024)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | β-adrenoceptor / NMR / Three dimentional structure / interaction / membrane / β作動薬 / 三次元構造 / リガンド-受容体相互作用 |
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| Research Abstract |
Based on the molecular modeling studies, the binding sites between these ligands (SWR-0342SA) and β-adrenoceptors (β-AR) were Asp104 and Leu335 in the β_1-receptors, and Asp117,Ser209,Leu303 and Ser192 and in β_3-adrenoceptors. Potencies of affinities of these compounds were also determined by the radioligand binding assay method. In addition, we analyzed the interaction sites between these compounds and β-AR by NMR. The sequence of 3^<rd> TMD peptide of the β_1- adrenergic receptor was found KKKELWTSVDVLAVTASIETLAVIALDRYLAKK. The peptide was synthesized and purchased from Sigma Genosys and purified to >74% purity as judged by HPLC. The purified peptide was prepared for NMR sample solution by dissolving it in DMSO solution. 1D HNMR and 2D NMR experiment were done in the JEOL 500MHz NMR machine. In 1D HNMR experiment presaturation method was done, whereas 2DNMR experiment was included DQFCOSY (double quantum filtered correlation spectroscopy), NOESY (Neclear Overhauser effects), ROESY (Rotating frame Overhauser spectroscopy), HMQC (hetaronuclear Multiple Quantum coherence) and HMBC (Hetaronuclear Multiple Bond Correlation). The ^1H NMR spectroscopy was performed where x_offset and x_sweep was 5.8 ppm and 12 ppm respectively. The ^<13>C NMR spectroscopy was also done where x_point was 95 ppm and x_sweep was 200 ppm. In NOESY experiment mixing time was 1.872 sec. The NMR spectra of bopindolol and 33 amino acid peptide of 3^<rd> TMD of β_1-AR were obtained individually. Finally we want to perform NMR analysis of complex between bopindolol and 3^<rd> TMD peptide of β_1-AR. We may be able to get the conformational structure of 33^<rd> TMD peptide from the bound and unbound NMR spectroscopy with bopindolol.
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Report
(3 results)
Research Products
(18 results)
