容量性カルシウム流入機構に関与する分子の生化学的解析

参与电容性钙内流机制的分子的生化分析

• 批准号: 14780588
• 负责人: 岩崎 広英
• 金额: $ 2.18万
• 依托单位: Okazaki National Research Institutes
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14780588/
• 关键词: カルシウムイオン; 容量性カルシウム流入; イノシトール3リン酸; 2-aminoethoxydiphenyl borate; イオンチャネル; 2-APB

細胞内においてカルシウムイオンはセカンドメッセンジャーとして機能し,多彩な生命現象に関与する.細胞質のカルシウムイオン濃度は平常時には低く抑えられており,刺激に応答して細胞内カルシウムストアからの放出または細胞外からの流入が生じる.本研究課題では,細胞内ストアからのカルシウムイオン放出に関与するイノシトール1,4,5-三リン酸受容体(IP3R)の阻害剤としてこれまで知られていた2-aminoethoxydiphenyl borate (2-APB)が,細胞内ストアの枯渇に伴う細胞外からのカルシウム流入,いわゆる容量性カルシウム流入(SOC)をも阻害することに着目し,その作用について解析することで容量性カルシウム流入の分子機構に迫ることを目的として実施された.容量性カルシウム流入に関与する形質膜上のチャネルの実体はこれまで明らかにされていない.2-APBはSOCに対するIC50値が5μMと高く,また特異性が低いため,結合分子の探索を試みることは不可能であった.そこで,2-APB類似化合物の中から2-APBよりも結合活性が高く,かつ特異性の高い化合物の探索を試みた.166個の2-APB類似化合物を探索した結果,数個の化合物が2-APBよりも強い阻害活性や特異性を有する化合物を見出すことに成功した.特にNo.163はSOCに対するIC50が59nMであり,2-APBよりも100倍も強い阻害活性を示したが,IP3Rに対する阻害効果も有していた(IC50は2-APBが20μMであるのに対しNo.163は10μM).これに対しNo.116は,SOCに対する阻害効果はNo.163より弱い(IC50=130nM)が,IP3Rに対する阻害効果を示さなかった.これらの化合物を見出したことにより,その結合タンパク質の解析が可能となり,生化学的解析が可能となった.現在これらの成果を投稿中である.

In the cell, there is a lot of energy in the cell, and there is a lot of energy in the cell, and there is a lot of color in life. In normal times, when the temperature is low, the cell temperature is low and the temperature is low, so as to stimulate the response of the cell to the cell. The extracellular signal is released from the cell and flows into the plant. In this study, we found that there was no significant difference between the two groups in this study. in this study, we found that there was no significant difference between the two groups in this study. in this study, the results of this study were as follows: in this study, we found that there was no significant difference between the two groups in this study. in this study, the results of this study were as follows: (1) in this study, the results of this study were as follows: (1) in this study, the results of this study were as follows: in this study, the intracellular and extracellular acid receptors (IP3R) were released from the cells in this study. in this study, we did not know that there was an influx of acid receptors in the cells. On the basis of the information on the flow of capacity into the market (SOC), the effect on the flow of capacity into the molecular mechanism is not required. The flow rate of capacitive particles into the film is different from that of the film. The volume of the IC50 is 5 μ M high, the specific low temperature is low, and the molecular probe is not possible. It is not possible to use the molecular method to explore the characteristics of the SOC. The 2-APB class is similar to the compound 2-APB complex with high activity, and the specific compound with high activity. 166 compounds with 2-APB class are similar to the results of the compound exploration test, and several compounds with 2-APB strong inhibitory activity have been successfully screened. Special No.163, SOC, 59nM, 2-APB, IP3R, APB, 20 μ M, 10 μ M, 163mm, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum, 10mum. The No.116, the SOC, the IC50=130nM, the IP3R, the weak, the weak. It is possible to analyze the compounds in combination with the chemical compounds, and to analyze the biochemical properties of the compounds. I am now in the process of submitting contributions to the achievements.