Generally, CPPs are defined as short, water soluble and partly hydrophobic and/or polybasic peptides (at most 30–35 amino acid residues) with a net positive charge at physiological pH [1]. This new class of peptides was introduced in the late 1980s by the discovery of the human immunodeficiency virus type 1 (HIV-1) encoded Tat peptide [2, 3] and the amphiphilic Drosophila Antennapedia homeodomain-derived 16 amino acid penetratin peptide (pAntp), which was discovered somewhat later [4–7]. These two peptides are the most extensively studied

of all CPPs. The main feature of CPPs is that they are able to penetrate the cell membrane at low micromolar concentrations in Inhibitors,research,lifescience,medical vivo and in vitro without using any chiral receptors and without causing irreversible membrane damage. These peptides are capable of internalizing electrostatically or covalently bound biologically active cargoes such as drugs, Inhibitors,research,lifescience,medical with high efficiency and

low toxicity [1, 8]. Despite many studies made on CPPs, the mechanism(s) by which CPPs enter the cells has not been completely resolved. There is Inhibitors,research,lifescience,medical some evidence for both energy-independent processes and endocytosis in internalization of CPPs. Presently, endocytosis, composed of two steps, endocytotic entry followed by endosomal escape, is believed to be the most common uptake mechanism at low CPP concentrations [8, 9]. Model biomembranes or lipid bilayers are efficient model systems to investigate the CPPs translocation mechanism(s). Large unilamellar vesicles (LUVs) are among the most commonly used model membranes in lipid-peptide interaction studies [10]. Here, we have performed experiments to study Inhibitors,research,lifescience,medical the background Inhibitors,research,lifescience,medical mechanism(s)

of endosomal escape. Cell membranes are normally weakly negatively charged and consist of different phospholipid molecules and associated proteins and proteoglycans. The lipids used in our study (a mixture of zwitterionic POPC and negatively charged POPG phospholipids) have been chosen to mimic cell membranes. Bacteriorhodopsin (BR) reconstituted Anacetrapib into LUVs with 20% negatively charged phospholipid are used to model the endosomes. The LUVs were prepared by the extrusion method, and their size and stability were carefully examined by dynamic light scattering (DLS). BR is an integral membrane protein of about 26KDa found in Halobacterium salinarium. There are various methods to reconstitute membrane proteins into the vesicles including organic solvent-mediated reconstitution, direct incorporation into preformed liposomes, mechanical means, and the detergent-mediated selleckchem Tipifarnib reconstitution method. Among these methods, detergent-mediated reconstitution is the most common and successful technique to incorporate membrane proteins into vesicles [11].