Chen Liping, Zhou Jun, Zhang Yunyun, Chu Shuangshuang, He Weina, Li Ye, Su Xiurong
School of Marine Sciences, Ningbo University, Ningbo, Zhejiang Province, People's Republic of China.
PLoS One. 2015 Apr 16;10(4):e0119427. doi: 10.1371/journal.pone.0119427. eCollection 2015.
Ferritin has important functions in the transition and storage of toxic metal ions, but its regulation and function in many invertebrate species are still largely unknown. In our previous work, the cDNA sequence of Sinonovacula constricta, Apostichopus japonicas and Acaudina leucoprocta were constructed and efficiently expressed in E. Coli BL21 under IPTG induction. In this follow-up study, the recombinant ferritins were exposed to heavy metal manganese. The manganese concentration levels in three recombinant ferritins were greater than horse spleen ferritin (HSF). Compared with HSF, the amount of manganese enrichment in the three recombinant ferritins was 1.75-fold, 3.25-fold and 2.42-fold increases in ScFER, AjFER, and AlFER, respectively. After phosphate stimulation, the concentration of manganese increased and was higher than the ordinary dialysis control groups. The ScFER was four times its baseline value. The AjFER and AlFER were 1.4- and 8-fold higher, respectively. The AlFER sample stimulated by phosphate was 22-fold that of HSF. The morphologies of the resulting Mn-Ferritin from different marine invertebrates were characterized with scanning electron microscopy. Surface morphologies were lamella flower-like and are consistent with changes in surface morphologies of the standard Mn-HSF. Invertebrate recombinant ferritin and HSF both can uptake manganese. We found that the structure of A. leucoproctarecombinant Mn-Ferritin aggregate changed over time. The surface formed lamella flower-like aggregate, but gradually merged to create a relatively uniform plate-like phase of aggregate spherically and fused without clear boundaries.
铁蛋白在有毒金属离子的转化和储存中具有重要功能，但在许多无脊椎动物物种中其调节和功能仍 largely 未知。在我们之前的工作中，构建了缢蛏、刺参和白氏海豆芽的 cDNA 序列，并在 IPTG 诱导下在大肠杆菌 BL21 中高效表达。在这项后续研究中，将重组铁蛋白暴露于重金属锰。三种重组铁蛋白中的锰浓度水平高于马脾铁蛋白（HSF）。与 HSF 相比，三种重组铁蛋白中锰的富集量在 ScFER、AjFER 和 AlFER 中分别增加了 1.75 倍、3.25 倍和 2.42 倍。磷酸盐刺激后，锰的浓度增加且高于普通透析对照组。ScFER 是其基线值的四倍。AjFER 和 AlFER 分别高出 1.4 倍和 8 倍。经磷酸盐刺激的 AlFER样品是 HSF 的 22 倍。用扫描电子显微镜对来自不同海洋无脊椎动物的所得锰铁蛋白的形态进行了表征。表面形态为片状花状，与标准锰 - HSF 的表面形态变化一致。无脊椎动物重组铁蛋白和 HSF 都能摄取锰。我们发现白氏海豆芽重组锰铁蛋白聚集体的结构随时间发生变化。表面形成片状花状聚集体，但逐渐融合形成相对均匀且无明显边界融合的球状板状聚集体相。
