Transduction of Proteins into Leishmania Tarentolae by Formation of Non-Covalent Complexes With Cell-Penetrating Peptides
Transduction of Proteins into Leishmania Tarentolae by Formation of Non-Covalent Complexes With Cell-Penetrating Peptides
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DOI：
10.1002/jcb.24654
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发表时间：
2014-02-01
影响因子：
4
通讯作者：
Reissmann, Siegmund
中科院分区：
文献类型：
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作者：
Keller, Andrea-Anneliese;Breitling, Reinhard;Reissmann, Siegmund
细胞穿透肽（CPP）用于将肽、蛋白质、不同类型的核糖核酸（或这些分子的模拟物）和DNA转运到植物和哺乳动物的活细胞中。与哺乳动物细胞相比，利什曼原虫属于具有不同的膜脂质组成、表面上的蛋白聚糖和信号通路的原生动物类。我们研究了两种不同的，容易检测到的蛋白质的非致病性菌株利什曼原虫tarentolae的摄取。从大量可用的CPP中，专门选择了六个和一个组蛋白，因为它们能够形成非共价复合物。对于利什曼原虫，我们使用半乳糖苷酶和荧光标记的牛血清白蛋白作为货物。将结果与使用哺乳动物细胞的类似内化研究进行比较[Mussbach等人，].利什曼原虫细胞可以通过分泌的膜结合胰凝乳蛋白酶样蛋白酶降解CPP。两种货物蛋白质都以足够的效率内化，并达到与哺乳动物细胞相似的分子内浓度。的CPP的运输效率彼此不同，并显示出不同的排名顺序为两种货物。荧光标记的牛血清白蛋白的细胞内分布显示在细胞核和动基体中的浓度最高。利什曼原虫对高浓度的一些CPP敏感，尽管与哺乳动物细胞不同。MPG-肽在利什曼原虫中比在哺乳动物细胞中更具细胞毒性，充当抗微生物肽。我们的研究结果有助于更好地了解利什曼原虫细胞中的分子相互作用，并可能对利什曼病的新治疗方法。J.细胞。115：243-252，2014. (c)2013 Wiley Periodicals，Inc.
Cell-penetrating peptides (CPPs) are used to transport peptides, proteins, different types of ribonucleic acids (or mimics of these molecules), and DNA into live cells, both plant and mammalian. Leishmania belongs to the class of protozoa having, in comparison to mammalian cells, a different lipid composition of the membrane, proteoglycans on the surface, and signal pathways. We investigated the uptake of two different and easily detectable proteins into the non-pathogenic strain Leishmania tarentolae. From the large number of CPPs available, six and a histone were chosen specifically for their ability to form non-covalent complexes. For Leishmania we used the enzyme -galactosidase and fluorescent labeled bovine serum albumin as cargoes. The results are compared to similar internalization studies using mammalian cells [Mussbach et al., ]. Leishmania cells can degrade CPPs by a secreted and membrane-bound chymotrypsin-like protease. Both cargo proteins were internalized with sufficient efficiency and achieved intramolecular concentrations similar to mammalian cells. The transport efficiencies of the CPPs differed from each other, and showed a different rank order for both cargoes. The intracellular distribution of fluorescent-labeled bovine serum albumin showed highest concentrations in the nucleus and kinetoplast. Leishmania are susceptible to high concentrations of some CPPs, although comparably dissimilar to mammalian cells. MPG-peptides are more cytotoxic in Leishmania than in mammalian cells, acting as antimicrobial peptides. Our results contribute to a better understanding of molecular interactions in Leishmania cells and possibly to new treatments of leishmaniasis. J. Cell. Biochem. 115: 243-252, 2014. (c) 2013 Wiley Periodicals, Inc.