CAJ | 학술논문

利用[~3H]-ryanodine结合实验,SDS-PAGE和Western Blotting,光子相干光谱法(PCS)和DPH荧光偏振法,考察氧化胁迫条件下氧化型通道调控剂1,4NQ和Na_2SeO_3对RyR1通道活性,SR膜蛋白分布,RyR1的平均粒径和SR膜流动性的影响.结果显示,高浓度的1,4NQ和Na_2SeO_3处理使RyR1通道活性和SR膜的流动性降低,并且导致SR上的膜蛋白交联形成大分子交联复合物,而RyR1参与了它的形成,DTT可以逆转交联复合物的的形成.结果提示,高浓度氧化剂对RyR1通道的抑制作用,可能是由于氧化了负责关闭通道的职能巯基导致蛋白间错误交联,从而影响了钙释放通道和钙释放单元的结构和功能.
이용[~3H]-ryanodine결합실험,SDS-PAGE화Western Blotting,광자상간광보법(PCS)화DPH형광편진법,고찰양화협박조건하양화형통도조공제1,4NQ화Na_2SeO_3대RyR1통도활성,SR막단백분포,RyR1적평균립경화SR막류동성적영향.결과현시,고농도적1,4NQ화Na_2SeO_3처리사RyR1통도활성화SR막적류동성강저,병차도치SR상적막단백교련형성대분자교련복합물,이RyR1삼여료타적형성,DTT가이역전교련복합물적적형성.결과제시,고농도양화제대RyR1통도적억제작용,가능시유우양화료부책관폐통도적직능구기도치단백간착오교련,종이영향료개석방통도화개석방단원적결구화공능.
By using [~3H]-ryanodine binding assay, SDS-PAGE, Western Blotting, photon correlation spectroscopy (PCS) and DPH fluorescence polarization, the influences of oxidation modulators 1,4NQ and Na_2SeO_3 on the channel activity, the average particle size of RyR1, the distribution of SR proteins in cross-linking complex, and fluidity of SR membrane under the oxidative stress were investigated.The results indicate that, upon to the oxidants treatment of 1,4NQ and Na_2SeO, both the activity of RyR1 channel and the fluidity of SR membrane decreased, and macromolecular cross-linked complexes consisting of RyR1 emerged on the gel of the SR membrane proteins.Further investigations showed that DTT decomposed the cross-linked complexes.Above results suggest that the inhibition of RyR1 channel caused by the high concentration of oxidant modulators is probably due to oxidation of the functional sulfhydryls which are responsible for the closure of the channels, and the occurrence of mistaken cross-linking between SR proteins which would alter the function of the calcium release unit.