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Kinetic analysis of the activation of photoactivated adenylyl cyclase (PAC), a blue-light receptor for photomovements of Euglena

机译:蓝藻光激活蓝光受体光活化腺苷酸环化酶(PAC)活化的动力学分析

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摘要

Photoactivated adenylyl cyclase (PAC) was first purified from a photosensing organelle (the paraflagellar body) of the unicellular flagellate Euglena gracilis, and is regarded as the photoreceptor for the step-up photophobic response. Here, we report the kinetic properties of photoactivation of PAC and a change in intracellular cAMP levels upon blue light irradiation. Activation of PAC was dependent both on photon fluence rate and duration of irradiation, between which reciprocity held well in the range of 2-50 μmol m-2 s-1 (total fluence of 1200 μmol m-2). Intermittent irradiation also caused activation of PAC in a photon fluence-dependent manner irrespective of cycle periods. Wavelength dependency of PAC activation showed prominent peaks in the UV-B/C, UV-A and blue regions of the spectrum. The time course of the changes in intracellular cAMP levels corresponded well with that of the step-up photophobic response. From this and the kinetic properties of PAC photoactivation, we concluded that an increase in intracellular cAMP levels evoked by photoactivation of PAC is a key event of the step-up photophobic response.
机译:光活化的腺苷酸环化酶(PAC)首先从单细胞鞭毛的裸藻Euglena gracilis的光敏细胞器(鞭毛旁体)中纯化,并被认为是逐步增强的疏水反应的光感受器。在这里,我们报告了P​​AC的光活化的动力学特性以及蓝光照射后细胞内cAMP水平的变化。 PAC的活化取决于光子通量率和辐照时间,两者之间的互易性在2-50μmolm-2 s-1(总通量为1200μmolm-2)的范围内保持良好。间歇照射也导致PAC活化,其光子注量依赖方式与周期无关。 PAC活化的波长依赖性在光谱的UV-B / C,UV-A和蓝色区域中显示出明显的峰。细胞内cAMP水平变化的时间过程与逐步增强的光反应的时间过程非常吻合。从这一点以及PAC光活化的动力学特性,我们得出结论,PAC的光活化引起的细胞内cAMP水平的升高是逐步提高光反应性的关键事件。

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