Laboratório Integrado de Bioquímica-Hatisaburo Masuda, UFRJ, Polo Barreto, Av. São José do Barreto nº 764, São Jose do Barreto, CEP 27971-550 Macaé, RJ, Brazil.
Int J Biol Sci. 2013 Aug 27;9(8):842-52. doi: 10.7150/ijbs.6628. eCollection 2013.
The physiological roles of polyphosphates (poly P) recently found in arthropod mitochondria remain obscure. Here, the possible involvement of poly P with reactive oxygen species generation in mitochondria of Rhipicephalus microplus embryos was investigated. Mitochondrial hexokinase and scavenger antioxidant enzymes, such as superoxide dismutase, catalase, and glutathione reductase were assayed during embryogenesis of R. microplus. The influence of poly P₃ and poly P₁₅ were analyzed during the period of higher enzymatic activity during embryogenesis. Both poly Ps inhibited hexokinase activity by up to 90% and, interestingly, the mitochondrial membrane exopolyphosphatase activity was stimulated by the hexokinase reaction product, glucose-6-phosphate. Poly P increased hydrogen peroxide generation in mitochondria in a situation where mitochondrial hexokinase is also active. The superoxide dismutase, catalase and glutathione reductase activities were higher during embryo cellularization, at the end of embryogenesis and during embryo segmentation, respectively. All of the enzymes were stimulated by poly P₃. However, superoxide dismutase was not affected by poly P₁₅, catalase activity was stimulated only at high concentrations and glutathione reductase was the only enzyme that was stimulated in the same way by both poly Ps. Altogether, our results indicate that inorganic polyphosphate and mitochondrial membrane exopolyphosphatase regulation can be correlated with the generation of reactive oxygen species in the mitochondria of R. microplus embryos.
多聚磷酸盐(poly P)在节肢动物线粒体中的生理作用最近才被发现,目前还不清楚。本研究探讨了多聚磷酸盐是否与 Rhipicephalus microplus 胚胎线粒体中活性氧的产生有关。在 R. microplus 胚胎发生过程中测定了线粒体己糖激酶和清除抗氧化酶(如超氧化物歧化酶、过氧化氢酶和谷胱甘肽还原酶)的活性。分析了 poly P₃ 和 poly P₁₅ 在胚胎发生过程中酶活性较高时期的影响。这两种多聚磷酸盐均可使己糖激酶活性降低高达 90%,有趣的是,葡萄糖-6-磷酸是通过己糖激酶反应产物刺激线粒体膜外多聚磷酸酶的活性。在存在线粒体己糖激酶活性的情况下,多聚磷酸盐会增加线粒体中过氧化氢的生成。超氧化物歧化酶、过氧化氢酶和谷胱甘肽还原酶的活性在胚胎细胞化、胚胎发生末期和胚胎分割期分别较高。所有的酶都被 poly P₃ 所刺激。然而,poly P₁₅ 对超氧化物歧化酶没有影响,只有在高浓度时,过氧化氢酶的活性才会被刺激,而谷胱甘肽还原酶是唯一一种被这两种多聚磷酸盐以相同方式刺激的酶。总之,我们的研究结果表明,无机多聚磷酸盐和线粒体膜外多聚磷酸酶的调节可能与 R. microplus 胚胎线粒体中活性氧的产生有关。