Department of Biology and CESAM, University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal,
Ecotoxicology. 2014 Mar;23(2):285-92. doi: 10.1007/s10646-013-1172-8. Epub 2014 Jan 8.
Phytochelatins (PCs) are thiol-rich peptides, enzymatically synthesized by plants and algae under exposure to certain metals (Cd, Pb, Zn, Ag, As, Cu). Due to their ability to bind metal ions, they play an important role in the cellular detoxification, forming stable metal-PC complexes that minimize the intracellular deleterious effects of metals. The aim of the present work was to evaluate the efficiency of PC-Cd chelation in the freshwater diatom Nitzschia palea under 0, 0.1 and 0.2 mg Cd L(-1), which induced different levels of oxidative stress. This objective was accomplished by the isolation of PC-Cd complexes through size exclusion chromatography. Two peaks were identified, corresponding to high molecular weight (HMW) and low molecular weight (LMW) complexes. In each of the complexes, Cd was quantified by inductively coupled plasma-mass spectrometry, thiol composition was determined by HPLC analysis and the efficiency of Cd chelation calculated by -SH/Cd ratios in HMW and LMW complexes at both Cd concentrations. Results showed that the majority of intracellular Cd was complexed with PCs (75.2-91.2 %). PCs-binding efficiency in this diatom species was higher at HMW than at LMW complexes and enhanced with the increase of Cd concentration exposure. Our work evidenced the important role of PCs as the main intracellular tolerance mechanism in this species. The efficiency increase of Cd-PC binding is related to the increment of PCs synthesis and to the number of Cd ions coordinately bonded to -SH groups in LMW and HMW complexes.
植物螯合肽(PCs)是富含巯基的肽,在暴露于某些金属(Cd、Pb、Zn、Ag、As、Cu)时,植物和藻类通过酶合成。由于它们能够结合金属离子,因此在细胞解毒中发挥着重要作用,形成稳定的金属-PC 配合物,最大限度地减少金属在细胞内的有害影响。本研究的目的是评估 PC-Cd 螯合在淡水硅藻 palea 中的效率,在 0、0.1 和 0.2mgCdL(-1) 下,诱导不同水平的氧化应激。这一目标是通过尺寸排阻色谱法分离 PC-Cd 配合物来实现的。鉴定出两个峰,分别对应高分子量(HMW)和低分子量(LMW)配合物。在每个配合物中,通过电感耦合等离子体质谱法定量 Cd,通过 HPLC 分析确定巯基组成,并根据 HMW 和 LMW 配合物中-Cd 比计算 Cd 螯合的效率在两个 Cd 浓度下。结果表明,大部分细胞内 Cd 与 PCs 结合(75.2-91.2%)。在该硅藻物种中,HMW 比 LMW 配合物的 PC 结合效率更高,并且随着 Cd 浓度暴露的增加而增强。我们的工作证明了 PCs 作为该物种主要细胞内耐受机制的重要作用。Cd-PC 结合效率的增加与 PCs 合成的增加以及 LMW 和 HMW 配合物中-Cd 离子与 -SH 基团配位的数量有关。