Henriques Sónia Troeira, Melo Manuel Nuno, Castanho Miguel A R B
Centro de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Lisbon, Portugal.
Mol Membr Biol. 2007 May-Jun;24(3):173-84. doi: 10.1080/09687860601102476.
Membrane translocation is a crucial issue when addressing the activity of both cell-penetrating and antimicrobial peptides. Translocation is responsible for the therapeutic potential of cell-penetrating peptides as drug carriers and can dictate the killing mechanisms, selectivity and efficiency of antimicrobial peptides. It is essential to evaluate if the internalization of cell-penetrating peptides is mediated by endocytosis and if it is able to internalize attached cargoes. The mode of action of an antimicrobial peptide cannot be fully understood if it is not known whether the peptide acts exclusively at the membrane level or also at the cytoplasm. Therefore, experimental methods to evaluate and quantify translocation processes are of first importance. In this work, over 20 methods described in the literature for the assessment of peptide translocation in vivo and in vitro, with and without attached macromolecular cargoes, are discussed and their applicability, advantages and disadvantages reviewed. In addition, a classification of these methods is proposed, based on common approaches to detect translocation.
在探讨细胞穿透肽和抗菌肽的活性时，膜转位是一个关键问题。转位决定了细胞穿透肽作为药物载体的治疗潜力，并且能够决定抗菌肽的杀伤机制、选择性和效率。评估细胞穿透肽的内化是否由内吞作用介导，以及它是否能够内化附着的货物至关重要。如果不知道抗菌肽是仅在膜水平起作用还是也在细胞质中起作用，就无法完全理解其作用模式。因此，评估和量化转位过程的实验方法至关重要。在这项工作中，我们讨论了文献中描述的20多种用于评估体内和体外肽转位的方法，包括有无附着大分子货物的情况，并对其适用性、优点和缺点进行了综述。此外，基于检测转位的常用方法，对这些方法进行了分类。
