Zahid Maliha, Robbins Paul D
Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Biomedical Science Tower, Pittsburgh, PA, USA.
Methods Mol Biol. 2011;683:277-89. doi: 10.1007/978-1-60761-919-2_20.
Protein transduction domains (PTD) or cell-penetrating peptides (CPPs) are small peptides that are able to carry proteins, nucleic acid, and particles across the cellular membranes into cells. PTDs can be classified into three types: (1) positively charged, cationic peptides, comprised of homopolymers of arginine, ornithine, or lysine; (2) hydrophobic peptides, derived from leader sequences of secreted proteins, and cell-type specific peptides; (3) tissue-specific, mainly amphipathic peptides identified by screening of peptide displaying phage libraries. The cationic and hydrophobic PTDs can efficiently transduce a variety of cell types in culture and in vivo, but in a nonspecific manner. In contrast, the tissue-specific transduction domains have more restricted transduction properties and presumably transduce cells through a different mechanism. In this chapter, we described methods for screening peptide phage display libraries for cell and tissue-specific transduction peptides both in cell culture and in vivo and for functional analysis of transduction.
蛋白质转导结构域（PTD）或细胞穿透肽（CPP）是能够携带蛋白质、核酸和颗粒穿过细胞膜进入细胞的小肽。PTD可分为三种类型：（1）带正电荷的阳离子肽，由精氨酸、鸟氨酸或赖氨酸的同聚物组成；（2）疏水肽，来源于分泌蛋白的前导序列，以及细胞类型特异性肽；（3）组织特异性肽，主要是通过筛选展示肽的噬菌体文库鉴定出的两亲性肽。阳离子和疏水PTD能够在体外培养和体内有效地转导多种细胞类型，但方式是非特异性的。相比之下，组织特异性转导结构域具有更受限的转导特性，推测是通过不同的机制转导细胞。在本章中，我们描述了在细胞培养和体内筛选肽噬菌体展示文库以获得细胞和组织特异性转导肽的方法，以及转导功能分析的方法。
