Tetrahymena thermophila granule lattice protein 3 improves solubility of sexual stage malaria antigens expressed in Escherichia coli.

Tetrahymena thermophila granule lattice protein 3 improves solubility of sexual stage malaria antigens expressed in Escherichia coli.
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DOI:
10.1016/j.pep.2022.106060
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发表时间:
2022-06
影响因子:
1.6
通讯作者:
Clark, Theodore G.
Clark, Theodore G.
中科院分区:
生物学4区
文献类型:
--
作者:
Akkale, Cengiz;Cassidy-Hanley, Donna Marie;Clark, Theodore G.

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对低成本生产的要求使细菌细胞成为生产疟疾重组亚单位疫苗的合理平台。然而,蛋白质的溶解度一直是原核表达系统的主要绊脚石。值得注意的例子包括传播阻断疫苗候选物Pfs 25和Pfs 48/45,它们在大肠杆菌中表达为重组蛋白时几乎完全不溶。各种溶解度标签已被用于有限的成功,在提高溶解度,虽然最近的研究与颗粒晶格蛋白1(Grl 1 p)从纤毛原生动物,嗜热四膜虫,已显示出希望。在这里,我们研究了一个相关的可溶性标签,颗粒晶格蛋白3(Grl 3 p)从T。将其与Grl 1 p和用于改善多个蛋白质靶标的溶解度的充分研究的麦芽糖结合蛋白(MBP)进行比较。我们发现Grl 3 p在与Pfs 25连接时表现为Grl 1 p,但在与Pfs 48/45配对时显着提高了溶解度。
The requirement for low cost manufacturing makes bacterial cells a logical platform for the production of recombinant subunit vaccines for malaria. However, protein solubility has been a major stumbling block with prokaryotic expression systems. Notable examples include the transmission blocking vaccine candidates, Pfs25 and Pfs48/45, which are almost entirely insoluble when expressed as recombinant proteins in Escherichia coli. Various solubility tags have been used with limited success in improving solubility, although recent studies with granule lattice protein 1 (Grl1p) from the ciliated protozoan, Tetrahymena thermophila, have shown promise. Here, we examine a related solubility tag, granule lattice protein 3 (Grl3p) from T. thermophila, and compare it to both Grl1p and the well-studied maltose binding protein (MBP) used to improve the solubility of multiple protein targets. We find that Grl3p performs comparably to Grl1p when linked to Pfs25 but significantly improves solubility when paired with Pfs48/45.
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