Dipartimento di Scienze Biochimiche A. Rossi Fanelli and Istituto Pasteur, Fondazione Cenci Bolognetti, Sapienza University of Rome, Rome, Italy.
Proteins. 2010 Feb 1;78(2):259-70. doi: 10.1002/prot.22536.
Oxidative stress is a widespread challenge for living organisms, and especially so for parasitic ones, given the fact that their hosts can produce reactive oxygen species (ROS) as a mechanism of defense. Thus, long lived parasites, such as the flatworm Schistosomes, have evolved refined enzymatic systems capable of detoxifying ROS. Among these, glutathione peroxidases (Gpx) are a family of sulfur or selenium-dependent isozymes sharing the ability to reduce peroxides using the reducing equivalents provided by glutathione or possibly small proteins such as thioredoxin. As for other frontline antioxidant enzymatic systems, Gpxs are localized in the tegument of the Schistosomes, the outermost defense layer. In this article, we present the first crystal structure at 1.0 and 1.7 A resolution of two recombinant SmGpxs, carrying the active site mutations Sec43Cys and Sec43Ser, respectively. The structures confirm that this enzyme belongs to the monomeric class 4 (phospholipid hydroperoxide) Gpx. In the case of the Sec to Cys mutant, the catalytic Cys residue is oxidized to sulfonic acid. By combining static crystallography with molecular dynamics simulations, we obtained insight into the substrate binding sites and the conformational changes relevant to catalysis, proposing a role for the unusual reactivity of the catalytic residue.
氧化应激是生物体普遍面临的挑战,对于寄生虫来说尤其如此,因为它们的宿主可以产生活性氧物种 (ROS) 作为防御机制。因此,像扁形虫血吸虫这样寿命长的寄生虫已经进化出了精细的酶系统,能够解毒 ROS。在这些酶中,谷胱甘肽过氧化物酶 (Gpx) 是一类硫或硒依赖性同工酶,它们具有使用谷胱甘肽或可能是小蛋白(如硫氧还蛋白)提供的还原当量还原过氧化物的能力。与其他一线抗氧化酶系统一样,Gpx 位于血吸虫的外膜——表皮中。在本文中,我们展示了两个重组 SmGpx 的晶体结构,分辨率分别为 1.0 和 1.7 A,这两个结构都带有活性位点突变 Sec43Cys 和 Sec43Ser。这些结构证实该酶属于单体类 4(磷脂氢过氧化物)Gpx。对于 Sec 到 Cys 突变体,催化 Cys 残基被氧化为磺酸。通过将静态晶体学与分子动力学模拟相结合,我们深入了解了与催化相关的底物结合位点和构象变化,提出了催化残基异常反应性的作用。