Liphard M, Bothe E, Schulte-Frohlinde D
Max-Planck Institut für Strahlenchemie, Mülheim a.d. Ruhr, FRG.
Int J Radiat Biol. 1990 Oct;58(4):589-602. doi: 10.1080/09553009014551951.
The yields of strand break formation (Gssb) in single-stranded DNA (ssDNA) initiated by radiation-generated OH radicals have been determined using the method of low-angle laser light scattering (LALLS). The irradiations were carried out in aqueous, N2O-saturated solutions in the absence and presence of oxygen and at different concentrations of glutathione (GSH). GSH exhibits a protective effect, which is shown to be mainly due to OH radical scavenging. To quantify this, the rate constants for the reactions of OH with GSH and DNA have been redetermined under our experimental conditions. The values obtained were 9.0 x 10(9) and 4.5 x 10(8) dm3 mol-1 s-1, respectively. From the Gssb values obtained under anoxic conditions it is concluded that GSH protects against strand breakage (in addition to OH scavenging) by reacting with DNA radicals in competition to strand break formation. The rate constant of the repair reaction is 8.1 x 10(4) dm3 mol-1 s-1 at room temperature. For irradiations carried out in the presence of oxygen the rate of strand break formation is determined by the decay of DNA peroxyl radicals. Under these conditions we observed no protective effect of GSH apart from OH radical scavenging. The results are compatible with those that are expected from the oxygen-fixation hypothesis.
采用低角度激光光散射(LALLS)方法测定了辐射产生的OH自由基引发的单链DNA(ssDNA)中链断裂形成的产率(Gssb)。辐照在无氧和有氧条件下的N₂O饱和水溶液中进行,并在不同谷胱甘肽(GSH)浓度下进行。GSH表现出保护作用,主要是由于其对OH自由基的清除作用。为了对此进行量化,在我们的实验条件下重新测定了OH与GSH和DNA反应的速率常数。得到的值分别为9.0×10⁹和4.5×10⁸ dm³ mol⁻¹ s⁻¹。从缺氧条件下获得的Gssb值可以得出结论,GSH通过与DNA自由基反应以竞争链断裂形成来防止链断裂(除了OH清除之外)。修复反应的速率常数在室温下为8.1×10⁴ dm³ mol⁻¹ s⁻¹。对于在有氧条件下进行的辐照,链断裂形成的速率由DNA过氧自由基的衰减决定。在这些条件下,我们观察到除了OH自由基清除之外,GSH没有保护作用。结果与氧固定假说预期的结果一致。